bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell type specificity on integrin α2 mediated functions in pancreatic cancer
Authors: Mia Md. S. , Sun Dali , Venkatachalem Sathish , Mathew Sijo 
Published: May 2022

BackgroundAmong the various cancers, pancreatic cancer has the third highest mortality rate per 100,000 population each year. In 2021, an estimated 60,430 people will be diagnosed with pancreatic cancer in the United States, with more than 48,220 deaths. Desmoplasia is a hallmark of pancreatic cancer. Cell surface receptor integrin promotes tumor progression by modulating the reciprocal signaling between the actin‐cytoskeleton and the desmoplastic tumor microenvironment. Integrin’s are heterodimeric cell surface receptors consisting of non‐covalently associated α and β subunits. Integrin β1 forms heterodimers with 12 different α subunits, and its expression is upregulated in pancreatic cancer. Integrin α2 is one of the α subunits that shows both pro and anti‐tumorigenic properties in cancer and belongs to the I domain group of integrin where the I domain in α subunit mediates the binding to matrix proteins. This study tested the hypothesis that in pancreatic cancer, upregulation of integrin β1 differentially regulates integrin α2 mediated cell functions in a cell‐specific manner.MethodTwo of the major pancreatic cancer types, Mia Paca‐2 and Panc‐1 cell lines were used in this study. The protein level of integrin β1 was downregulated using genetic approaches and confirmed by immunoblotting, immunocytochemistry, and qRT‐PCR. Cell spreading and adhesion assays were performed with WT cells and clones with lower integrin β1 expression. Cells were grown on the 3D matrix to see the ability of cells to proliferate in the presence of matrix proteins.ResultsDownregulation of integrin β1 in both Mia Paca‐2 and Panc‐1 cells decreased tumor growth in the collagen‐Matrigel 3D matrix significantly (n=10, p<0.0001). With the reduction of expression of integrin β1, there is a significant decrease in the expression of integrin α5 in Mia paca‐2 and Panc‐1 cell, which resulted in a significant decrease in cell adhesion (n=3; P<0.05) and spreading (n=40; p<0.05) on fibronectin and vitronectin. However, there were contrasting results in the case of integrin α2 expression and functions with integrin β1 downregulation. An increase in integrin α2 expression was recorded in Miapaca‐2 cells (n=3, p<0.05) with increased cell adhesion (n=3; p<0.05) and cell spreading (n=3; p<0.05) on collagen‐1 by the deletion of integrin β1. In the case of Panc‐1, there is reduced expression of integrin α2 that resulted in decreased cell adhesion (n=3; p<0.05) and cellular spreading (n=30; p<0.05) on collagen 1 with the knock‐down of integrin β1.ConclusionThe results indicate that the tumor suppressor property of integrin α2 varies between different tumor types in pancreatic cancer. Further studies are required to understand the mechanism of the anti‐tumorigenic property of integrin α2 in pancreatic cancer.
Does VEGF Expression Correlate with Lymphatic Metastasis in Breast Cancer?
Authors: Rukmini Ashita , Mathur Sandeep , Vuthaluru Seenu 
Published: May 2022

IntroductionPrimary tumor characteristics are increasingly used for prognostication, predicting lymph node metastasis and planning adjuvant treatment in breast cancer. Vascular Endothelial Growth factor (VEGF) has been postulated to stimulate lymph vessels development and growth in tumors. Few studies on cell lines have documented how the new lymphatics develop or tumor cells invade these lymphatics. However, in human breast cancer the literature is sparse. Hence, this study is done to see the expression of VEGF and its correlation with lymphatic invasion in breast cancer patients.Materials & MethodsFormalin fixed paraffin embedded blocks of breast cancer specimens and controls were retrieved & 4u thick sections stained with H&E stains for morphological evaluation. Serial sections were cut on APES (3‐Triethoxysilylpropylamine) coated slides for performing immunohistochemistry using monoclonal antibodies against VEGF and DII40. The expression intensity of VEGF and its distribution in the tumor samples were observed and semi‐quantitatively analyzed. VEGF expression was interpreted as follows: negative (0): <5% staining; weak (+): 5–25% staining; positive (++): 26–50% staining; strongly positive (+++): >50% staining. Statistical analysis: samples with 0/+ staining were considered in low VEGF expression, while samples with ++/+++ expression were considered as high VEGF expression. For DII40 well demarcated vessels showing positivity in endothelial cells were counted per 10hpf after selecting hot spots at scanner view. For statistical analysis, 0 was considered as no expression and 1/10HPF was considered as high expression. Association between levels of VEGF and D II 40 was examined by the two‐way scatter plot and a linear regression analysis of DII 40 over VEGF. The association of positivity between the two markers was assessed using Chi Square test.ResultsHigh VEGF expression was seen 33 of the 43 evaluable patients (76.74%%; 95% CI 61.37%‐88.24%) and high DII 40 expression was seen in 30 of the 44 patients (68.18%; 95% CI 52.42%‐81.39%). Of the 33 patients with high VEGF expression, DII 40 was expressed in 26 of them (78.79%). There was a strong association between VEGF expression and DII 40 expression (p<0.01) (Figure 1). The variance explained by VEGF level in the variance of DII 40 as measured by the correlation (R2) was 0.1265 (p=0.02). When the expression of VEGF and D II 40 was compared with tumor characteristics, there was statistically significant correlation with lymphovascular invasion (LVI) but not with tumor size, type, grade, ER/PR status, Her2neu status or lymph node involvement.ConclusionVEGF expression is high in most tumors. There is significant association between LVI & lymph vessels detection as studied by D II 40 expression and VEGF expression. Further studies in clinical settings are essential before incorporating VEGF expression for prognostication and customized patient care.
Parkinson's Disease
Authors: Bai Kathleen , Cai Ziming , Cherif Ibrahim , Compaore Faltas C. , Fujikawa Ayaka , Hao Delilah , Jia Rachel , Koziakova Sabina C. , Liu Lenore , Qiu Elaine , Ramirez Victoria , Christensen Anna , Zhang Josh , Greig James , MacGillivray Brendan C. 
Published: May 2022

Parkinson’s disease (PD) is a neurodegenerative disease that involves a progressive loss of dopaminergic neurons within the substantia nigra. Patients with PD experience a loss of motor function over time. In normal patients, muscle contractions are driven by action potentials through the binding of the neurotransmitter dopamine. Disruptions to dopaminergic activity would thus result in less efficient contractions. Patients with Parkinson’s can experience akinesia, tremors and other motor dysfunction.Glucocerebrosidase (GCase), specifically β‐GCase, is a lysosomal enzyme involved in sphingolipid metabolism. β‐GCase falls under the hydrolase classification, allowing it to catabolize glucosylceramide (GlcCer) into glucose and ceramide. β‐GCase consists of 497 amino acid glycoproteins within its four domains. The glucocerebrosidase gene (GBA) mutations are often associated with Gaucher disease. However, β‐GCase synthesized from mutated GBA1 is often studied as a biomarker for PD susceptibility. In a healthy cell with the non‐mutated GBA gene, the GBA gene would be transcribed into mRNA and then transported out to the rough Endoplasmic Reticulum, where GCase is then synthesized. Lysosomal integral membrane protein‐2 (LIMP2) is a protein that transports GCase through the Golgi Apparatus and transfers the β‐GCase into a late endosome. When the late endosome fuses with a lysosome, where β‐GCase can hydrolyze its substrates. Mutated β‐GCase may affect this autophagy pathway.This research will explore how mutated β‐GCase may disturb lysosomal functions, which would consequently lead to the aggregation of alpha‐synuclein (ASN), a protein that regulates synaptic vesicle trafficking and subsequently releases dopamine. Therefore, the detection and accumulation of mutated lysosomal enzymes, such as β‐GCase, in the cerebrospinal fluid might serve as biomarkers for Parkinson’s Disease.
Comparative Analysis of Annexin A1 (ANXA1) Protein Expression in Different Cancer Types
Authors: Olascuaga‐Castillo Karyn , Rubio‐Guevara Susana , Cáceres‐Andonaire Elena , Caballero‐Aquiño Olga , Hilario‐Vargas Julio , Altamirano‐Sarmiento Dan , Morgado‐Díaz José Andrés , Berezovski Maxim 
Published: May 2022

Annexin A1 (ANXA1) is the first member of the Lipocortin family, a calcium‐dependent phospholipid‐binding protein with potent immunomodulatory activity. ANXA1 is related to cellular proliferation, apoptosis, progression, and metastasis in cancer, and its expression is variable depending on the tumor type. Our research will compare the expression of ANXA1 in various forms of cancer and establish if the protein is up‐or down‐regulated.MethodsWe analyzed 1904 immunohistochemistry datasets from The Human Protein Atlas (HPA). The samples represent 20 different forms of cancer from 455 patients. The expression of ANXA1 was compared to the same in tissue samples from 137 healthy subjects, which meant the tissues of origins of primary tumors. The levels of ANXA1 expression were quantified using a visual grading method based on staining intensity (I) and cell fraction stained (F). The Q score (combination of stained cell fraction and staining intensity) was obtained by the product of (F)*(I).Results9 of 20 cancer types showed increased protein expression (up‐regulation) comparison to their healthy counterparts. Eight cancer types showed decreased protein expression (down‐regulation). Three types of cancer showed no significant differences.ConclusionANXA1 expression may serve as an important diagnostic marker for such a variety of different cancer types.
Differences in Cellular Metabolism and Metabolic Regulation between Non‐diabetic and Diabetic Human Coronary Artery Endothelial Cells
Authors: Xu Cynthia M. , Karbasiafshar Catherine , Braga Karla M. S. , Teixiera Rayane B. , Sellke Frank W. , Abid M. R. 
Published: May 2022

ObjectiveTo determine the differences in cellular metabolism and metabolic regulation in non‐diabetic and diabetic human coronary artery endothelial cells (HCAEC and DM‐HCAEC) to further the understanding of the effects of metabolic syndrome on endothelial dysfunction in the setting of cardiovascular disease.MethodsBoth HCAEC (n = 5) and DM‐HCAEC (n = 5) were cultured to passage 6. The cells were then starved for 18 hours. After treatment, a Reactive Oxygen Species (ROS) Assay was performed, and cell lysates were made for proteomics analysis and western blotting.ResultsThe ROS assay demonstrated that the total ROS was significantly higher in DM‐HCAEC (p<0.0001) than in HCAEC. Proteomics analysis and western blotting showed that DM‐HCAECs had significantly altered cellular transport, RNA processing, protein biogenesis, and antioxidant pathways. Proteomics analysis demonstrated that there were about 1,500 proteins significantly upregulated or downregulated in DM‐HCAEC compared to HCAEC. Analysis using Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed a significant increase in the “vesicle‐mediated transport” pathway and significant decrease in the “mRNA metabolic process” pathway in DM‐HCAEC with false discovery rates (FDR) of 1.40 x 10‐58 and 4.90 x 10‐92, respectively. Western blot analysis showed a significant changes in phosphorylated AMPK/total AMPK, phosphorylated mTOR (serine 2481), total mTOR, and several enzymes responsible for the regulation or elimination of ROS (Figure 1). The increased levels of AMPK activation and dysregulation of mTOR may be due to the excessive ROS in DM‐HCAEC, which are consistent with the ROS assay findings and significant changes in expression of enzymes that control ROS levels. Further research is needed to determine the mechanisms of downstream effects.ConclusionsAMPK and mTOR are key regulators of energy homeostasis, cellular growth and metabolism, and are altered in diabetes in the setting of increased oxidative stress. These findings will guide further experiments to assess the effect of metabolic syndrome and diabetes on therapeutics targeted at improving the outcomes of both acute and chronic myocardial ischemia. This also underlines the need for further development of therapies or the use of current therapies to address the metabolic disturbances found in diabetes to improve clinical outcomes in cardiovascular disease.
Liver Proteomic Profile in Mice on a High‐Fat Diet: Modulation with Anti‐Tumor Intervention
Authors: Harber Isabelle , Zedan Dania , McClintock Shannon , Varani James , Aslam Muhammad N. 
Published: May 2022

ntroductionNonalcoholic fatty liver disease (NAFLD) is becoming a primary cause of liver damage in Western society. Nonalcoholic steatohepatitis (NASH) is an advanced stage of NAFLD and may lead to liver cancer. Our previous long‐term murine studies have shown the beneficial role of Aquamin, a red marine algae‐derived (calcium, magnesium, additional trace element‐rich) supplement, in reducing liver injury and decreasing liver tumor incidence. The main objective of the study was to determine how manifestations and effects of NASH can be mitigated using Aquamin. We hypothesized that mice on a high‐fat Western diet (HFWD) with minerals would exhibit less liver injury than mice on a HFWD without minerals.MethodsTwo cohorts of MS‐NASH mice (8‐week‐old) were placed on a HFWD with and without minerals for 16 weeks. An additional cohort of MS‐NASH mice on the same HFWD was treated with obeticholic acid (OCA). C57BL/6 mice on a regular chow were included as a control. During the in‐life phase of the study, weight changes were assessed weekly. At termination, livers were histologically assessed for steatosis and fibrosis by using hematoxylin and eosin, and picrosirius red staining. Liver tissue samples were also subjected to tandem mass tag (TMT) mass‐spectroscopic proteomics for protein expression profile in individual mice.ResultsMice on HFWD gained more weight than mice on control diet, but there was no overall change in weight for mice on HFWD, irrespective of interventions. On histological assessment, there was no difference in steatosis between the two high‐fat groups, but mice on Aquamin showed reduced picrosirius red staining or collagen deposition as compared to placebo (on HFWD) mice. Regarding proteomic profile, there was a clear distinction among control and HFWD groups. Placebo mice were used for comparison. Aquamin intervention altered 91 proteins while OCA altered 84 proteins in liver samples with a 2‐fold change. Of these, 57 proteins were common between Aquamin and OCA. Most of the proteins were upregulated. Aquamin upregulated 39 unique pathways (with a p‐value <0.05) as assessed by the Reactome database. Some of the pathways significantly impacted by Aquamin were formation of cornified envelope; keratinization; gap junction assembly; type I hemidesmosome assembly; apoptosis‐related; plasma lipoprotein assembly, remodeling and clearance; and hedgehog “off” state pathways. Proteins downregulated by OCA are mainly involved in the synthesis of bile acids and bile salts.ConclusionThe addition of dietary minerals may play a protective role in interfering with downstream advancement from steatosis to NASH. Our studies provide mechanistic insight into how mineral supplementation may contribute to the reduction in liver injury and tumor formation (one of the most devastating consequences) of fatty liver disease in the context of HFWD‐induced steatosis.
ErbB3 Promotes Intestinal Barrier Function and Expression of the Tight Junctional Protein Pmp22
Authors: Hsieh Jonathan J. , Almohazey Dana , Frey Mark R. 
Published: May 2022

BackgroundErbB3 is a member of the ErbB/EGFR family of receptor tyrosine kinases, and is the constitutively‐expressed neuregulin (NRG) receptor in the intestinal epithelium. Pathways downstream of ErbB3 have been implicated in regulating tight junctions in other systems, but a role for ErbB3 in barrier function in the gut has not been described. Here we tested the hypothesis that ErbB3 is required for maintenance of normal barrier function in the intestine.MethodsWe generated Villin‐Cre;ErbB3flox/flox mice (ErbB3‐IEKO) with ErbB3 deletion in the intestinal epithelium. All comparisons are to ErbB3flox/flox littermate controls. Mice were given a FITC‐dextran 4 kDa gavage to assess intestinal permeability. Ileal samples were collected for RT‐qPCR, immunohistochemistry, and bulk RNA‐seq analysis. Enteroids were generated and treated with the ErbB3 ligand NRG‐1β. Caco‐2BBe cells were grown on Transwells, treated with NRG‐1β, and assessed for TEER and FITC‐dextran (4kDa) flux.ResultsErbB3‐IEKO mice displayed increased intestinal barrier permeability to 4kDa FITC‐dextran compared to controls (p < 0.01). RNA‐seq analysis showed a decrease in the expression of the putative tight junction protein Pmp22in ErbB3‐IEKO mice, while other tight junctional components were unchanged. RT‐qPCR analysis of Ileal samples showed an 80% Pmp22downregulation with ErbB3 loss (p < 0.01) compared to controls, with similar results in ErbB3‐deficient enteroids. Immunohistochemical analysis confirmed near complete loss of PMP22 in ErbB3‐IEKO mice. In vitro, treatment with NRG‐1β induced Pmp22expression in control enteroids (p < 0.001) but not in ErbB3‐deficient enteroids. Furthermore, treatment of Caco‐2BBe monolayers with NRG‐1β increased TEER and reduced FITC‐dextran flux, demonstrating active regulation of permeability.ConclusionsWe demonstrate that loss of ErbB3 in the intestinal epithelium leads to reduced expression of the tight junction protein PMP22 and impaired barrier function in mice. This may represent a mechanism for regulation of the intestinal barrier through ErbB3 and PMP22. Furthermore, increased TEER in Caco‐2BBe monolayers following NRG‐1β treatment suggests induction of PMP22 may represent a means to enhance barrier function. Since disrupted intestinal barrier function contributes to the pathophysiology of chronic inflammatory conditions such as inflammatory bowel disease, these results may point to potential future therapeutic interventions targeting the barrier.
Phosphoglycerate Mutase 5 Expression Modulates Hepatocellular Steatosis
Authors: Johnston Andrea , Moeller Cambri , Liu Chin‐Chi 
Published: May 2022

Phosphoglycerate Mutase 5 (PGAM5) is a serine/threonine phosphatase that plays a role in oxidant injury sensing, mitochondrial biogenesis, mitophagy, and multiple cell death pathways.Pgam5 null mice are protected from high‐fat diet induced obesity. This metabolic phenotype was attributed to upregulation of adaptive thermogenesis in brown adipose tissue, but hepatocyte specific lipid metabolism has not been evaluated. We hypothesize that hepatic PGAM5 expression level modulates hepatic steatosis. Using primary cultures of Pgam5 knockout hepatocytes, we have shown that palmitate induced steatosis is attenuated and that treatment with the long‐chain fatty acid impairs oxygen consumption rate specifically reducing the maximal respiratory capacity. Further, over‐expression of PGAM5 in human hepatoma cells exacerbates palmitate induced steatosis. Cumulatively, these results suggest that PGAM5 regulates hepatic lipogenesis and mitochondrial long‐chain fatty acid beta‐oxidation.
Nitazenes are potent mu‐opioid receptor agonists with profound respiratory depression
Authors: Palkovic Barbara , Malcom Nicholas J. , McCorvy John D. , Langer Thomas M. , Callison Jennifer J. , Stuth Eckehard A. , Zuperku Edward J. , Stucke Astrid G. 
Published: May 2022

BackgroundNitazene opioids (2‐benzylbenzimidazole opioids) were first identified in multiple overdose fatalities a few years ago. It has been shown that nitazene drugs are highly active at the mu‐opioid receptor, with potency and efficacy exceeding fentanyl. Because of this, isotonitazene was added to Schedule I of the controlled substance list in August 2020 by the Drug Enforcement Agency. Further studies have shown the high potency of the N‐desethyl‐isotonitazene metabolite, which was similar in potency of isotonitazene itself. The objective of this study was to compare the pharmacokinetics with regards to respiratory depression between N‐desethyl‐isotonitazene and fentanyl in vivo rabbit model.MethodsThe study was approved by the local Animal Care Committee and conformed to NIH standards. Adult New Zealand White rabbits (3‐4kg) were anesthetized, tracheotomized, ventilated, decerebrated and vagotomized. Phrenic nerve activity was recorded from the c5 rootlet. Phrenic neural activity, respiratory rate, arterial blood pressure, and airway carbon dioxide concentration were recorded on a computerized chart recorder (Powerlab/16SP; ADInstrumentes, Australia). We determined the magnitude and time course of respiratory rate depression from 1mcg/kg N‐desethyl‐isotonitazene and fentanyl and subsequently the dose required to achieve apnea. Data were collected using LabChart data and exported to SigmaPlot 11 (Systat Software, USA) for data processing, data plotting and statistical analysis. Using the phrenic neurogram, we determined the respiratory rate and peak phrenic activity. We plotted the average respiratory rate for each minute normalized to control frequency, removed artefacts, and determined the best fitting curve for each plot. Plots were averaged for each dose and displayed as mean± standard error.ResultsFentanyl (1mcg/kg) depressed respiratory rate by 25±5% from baseline (n=3) while N‐desethyl‐isotonitazene caused a 38±2% depression (n=6). Recovery to baseline was 150±15 minutes for N‐desethyl‐isotonitazene and 46±8 minutes for fentanyl. N‐desethyl‐isotonitazene caused apnea at 4mcg/kg (n=6), and complete recovery to baseline respiratory rate was 201±33 min minutes. Fentanyl caused apnea at 8mcg/kg (n=4), and 64±7 minutes were needed to recovery.ConclusionsNitazenes are potent synthetic opioids. At equal doses, they cause more profound and longer lasting respiratory rate depression than fentanyl.
H2S Therapy Improves Diastolic Dysfunction and Exercise Capacity in Two‐Hit Murine Model of Heart Failure with Preserved Ejection Fraction
Authors: Doiron Jake E. , Lapenna Kyle , Li Zhen , Xia Huijing , Goodchild Traci , Xian Ming , Kang Jianming , Lefer David 
Published: May 2022

BackgroundHFpEF is a complex multi‐organ disease expected to represent >60% of all clinical heart failure cases by 2030. To date, the majority of therapeutic approaches to treat HFpEF have largely failed in either preclinical or clinical trials. Given the recent reports of diminished H2S bioavailability in HFpEF combined with the robust cardioprotective actions of H2S in cardiovascular disease, we sought out to test the effects of H2S therapy in a well‐established pre‐clinical HFpEF murine model.MethodsHFpEF was induced in nine‐week‐old C57/BL6N mice (n=10 per group) by high fat diet and L‐NAME (0.5g/L/day) via drinking water for 5 weeks. Mice were treated with either vehicle or the H2S donor, JK‐1 (100 mg/kg/day B.I.D., I.P.) for a period of 5 weeks. Echocardiography and exercise capacity were performed at study weeks 0, 5, and 10 for monitoring of cardiac function and progression of exercise intolerance. At week 10, endothelium‐dependent and independent aortic vascular reactivity and left ventricular invasive hemodynamic studies were performed.ResultsTreatment with JK‐1 significantly improved cardiac function as measured by decreases in both left ventricular end diastolic pressure (LVEDP) and E/e’ after 5 weeks of treatment. Similarly, endothelium‐dependent vascular relaxation with acetylcholine exhibited significant improvement. Treadmill exercise performance expressed as total work showed improvement for the JK‐1 group, demonstrating changes in exercise capacity are irrespective of body weight.ConclusionThese results suggest that the H2S donation therapy exerts beneficial effects in the setting of severe HFpEF. Studies are currently ongoing to determine the mechanisms by which H2S therapy improves both cardiac and vascular function in this devastating cardiovascular disease.