bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Systematic Review of Assessment Practices for Self‐Directed Learning in Health Professions Education
Authors: Lerchenfeldt Sarah , Kemp Kyeorda , Mi Misa , Taylor Tracey A. 
Published: May 2022

IntroductionThis systematic review aimed to describe the assessment practices related to self‐directed learning (SDL) in undergraduate health professions education. There were three main objectives of the review: 1) evaluating the different types of assessment used to determine the effectiveness of SDL; 2) assessing the effectiveness of SDL methods on student learning; and 3) exploring how readiness for SDL is assessed.MethodsThe PRISMA standards were followed for this systematic review. A total of seven electronic databases were searched: CINAHL, Embase, ERIC, PsycINFO, PubMed, Scopus, and Web of Science. Articles published in English between 2015 and March 2020 were retrieved for further evaluation. Two independent investigators screened abstracts and full‐text articles using the inclusion and exclusion criteria. The same investigators independently extracted data from included articles: the type of participant, sample size, study design, SDL activities, and assessment methods used to evaluate effectiveness of SDL. In addition, two independent investigators evaluated study quality using the validated Medical Education Research Study Quality Instrument (MERSQI). A third investigator resolved discrepancies and disagreements.ResultsA total of 65 studies from nineteen different countries were included in the analysis. Most study participants were medical (N=33) or nursing (N=20) students. Overall, there were wide variations in the definition of SDL, SDL activities used, and in the assessments developed to evaluate the effectiveness of SDL activities. The most common type of assessment used was a student perception survey, unrelated to a SDL readiness assessment. Internally‐developed knowledge exams were also commonly used. A total of 38 studies concluded that the SDL had a positive outcome, measured by the assessment instrument, whereas 21 studies reported a mixed or neutral outcome and six studies reported a negative outcome.ConclusionsThis systematic review demonstrates the wide variations in both the SDL activities used as well as the assessment methods developed to determine SDL effectiveness. Since SDL is a necessary skill for lifelong learning and success as a healthcare professional, it must be used and assessed appropriately. It may be valuable to standardize SDL activities and assessment methods across healthcare professions education. In addition, future research should be conducted to help establish the most accurate and effective assessment methods for SDL.
Interleukin‐18 Enhances Contractile Responses to Phenylephrine in Pudendal Artery from Wistar Rats
Authors: Arishe Olufunke O. , Priviero Fernanda , Wilczynski Stephanie , Webb Clinton 
Published: May 2022

BackgroundHypertension is a major risk factor for the development of cardiovascular diseases and is a leading cause of mortality. The etiology of essential hypertension remains unclear, and its mechanisms are complex. Inflammation is a complex phenomenon, comprises the first response of the immune system in the presence of harmful stimuli and is considered to be a major cause of increased blood pressure. Indeed, low‐grade inflammation and activation of the immune system have been implicated in hypertension. Cytokine levels are elevated in hypertension and have been reported to be implicated in the associated disturbances in vascular reactivity reported in this disease. Clinical and population studies have consistently found increased circulating levels of interleukin 18 (IL‐18) in patients with hypertension.HypothesisThis study tests the hypothesis that IL‐18 will enhance contractile responses to phenylephrine in isolated arterial rings from rats.MethodsVascular function was performed on isolated pudendal arteries from male Wistar rats using wire myographs, kept in Krebs solution at 37°C, constantly aerated with 95%O2/5%CO2. Concentration‐response curves to phenylephrine (PE; 10‐9‐3x10‐5M), Acetylcholine (ACH; 10‐9 – 3x10‐5M) were performed in the absence or in the presence of IL‐18 (20ng/ml) for 1 hour. 10 minutes prior to the incubation with IL‐18, some arterial rings were incubated separately with the following agents: NADPH oxidase 2 inhibitor, gp91 ds‐tat (10‐5M); NADPH oxidase 1 and 4 inhibitor, GKT137831(10‐6M); and mitogen‐activated protein kinase (MKK / MEK) inhibitor, PD98059 (10‐5M) Concentration‐response curves were analyzed using non‐linear regression analysis. Data are presented as mean ± S.E.M. Statistical significance set at p<0.05. Data was analyzed with GraphPad Prism 9.2.0.ResultsIn the Wistar pudendal arterial rings, the potency of PE was increased in the presence of IL‐18 (pEC506.510 ± 0.3987 vs 6.022 ± 0.2738 for control; p = 0.0268) A, IL‐18 plus gp91 ds‐tat (pEC506.643 ± 0.2215 vs 6.264 ± 0.2471 for control; p = 0.0058) B and IL‐18 plus GKT137831 (pEC506.700 ± 0.2937 vs 6.272 ± 0.2963 for control; p = 0.0003) C. Incubation with IL‐18 plus PD98059 restored the contraction to PE to control levels. (pEC506.105 ± 0.4007 vs 6.170 ± 0.2847 for control; p = 0.7120) D. The relaxation response to ACh was not changed by IL‐18 after incubation for 1 hr.ConclusionOur results show that IL‐18 enhances the contractile responses to PE in pudendal arteries from Wistar rats by activation of the Mitogen‐Activated Protein Kinase/Extracellular Signal‐Regulated Kinase Pathway.
pHire: A Novel Genetically Encoded Red Intracellular pH Sensor Expressed in Cells and Flies
Authors: Schreiber Areonna C. , Rossano Adam J. , Reynolds Carmen J. , Holmes Heather L. , Brown Matthew R. , Matveyenko Aleksey , Romero Michael F. 
Published: May 2022

Intracellular pH regulation is vital for normal cellular function and is controlled by membrane H+ and HCO3‑ transporters. Most pH reporters have green fluorescence (i.e., BCECF, pHlorin) making simultaneous Ca2+, Cl‐, or other sensing (i.e., green fluorescent sensors) or GFP‐tags impossible. We previously reported a red‐fluorescent pH sensor (pHire) with an alkaline‐shifted pKa that increases fluorescence with increasing intracellular pH. pHire was transiently transfect into TM5 cells, but these fibroblast‐like cells did not address continuous expression in epithelial or excitable cells. Our objective was to evaluate pHire responses in multiple cell‐types and show that it can be used for fluorescent multiplexing.We selected pHire‐expressing rat pancreatic β‐cells (INS1pHire) and porcine kidney epithelial cells (LLCPK1pHire), and then FACS sorted to keep the top 10‐20% fluorescent cells. To test pHire’s usefulness for intact animals, we made transgenic, UAS‐pHire Drosophila, allowing pHire expression in specific tissues via genetic crossing with promoter‐GAL4 lines. By using perfusion chambers, we monitored cellular pHire‐responses to acid‐loading (NH4Cl pulse) and CO2/HCO3‑ addition.Stable selection of pHire cell lines ensures single cells as well as contiguous cell layers for acid‐base transporter evaluation. Addition of NH4Cl quickly alkalinizes all pHire cells. Different acid‐recovery rates were found in individual cells with LLCPK1pHire cells recovering faster than INS1pHire cells. CO2/HCO3‑ addition acidified both mammalian cell‐types. LLCPK1pHire cells typically show a Na+ dependent recovery (consistent with electrogenic Na+ bicarbonate cotransporter, NBCe1, activity) that is absent from INS1pHire cells. Infection of INS1pHire cells with a NBCe1A‐GFP AAV allowed clear demonstration of NBCe1 activity.To test pHire’s stability in living animals, we next generated a UAS‐pHire transgenic Drosophila line, allowing pHire expression in specific tissues via genetic crossing with various promoter‐GAL4 lines. We tested renal epithelial (principal cells: Uro‐GAL4, CapaR‐GAL4; stellate cells: c724‐GAL4) and CNS (elav‐GAL4) expression. As observed in mammalian cells, NH4Cl acid‐loading showed robust and more uniform non‐HCO3‑ recovery. The pH‐calibration curves for pHire in Drosophila cells were similar those obtained in mammalian cells (pKa>7.4). Thus, pHire characteristics are not changed by continuous expression in whole organisms.This study shows that the pHire sensor can be constitutively expressed into multiple cell lines or whole animals (flies) with an ideal pH range of pH 6 to 9. pHire may also be multiplexed with GFP labels and biosensors to allow real‐time, pH measurements with other ions of interest at a cellular scale. Finally, our data demonstrate that genetically encoded pHire is a reliable and useful tool for studying acid‐base transporters in a variety of cell‐types.
IL‐19 Regulates Atherosclerotic Plaque Progression via Lymphangiogenesis
Authors: Peluzzo Amanda , Preston Kyle , Leigh Tani , Kelemen Sheri , Autieri Michael 
Published: May 2022

IntroductionCardiovascular diseases are the leading cause of deaths globally, representing approximately 31% of deaths per year, 85% of which are due to myocardial infarction (MI) or stroke. This does not account for the millions of patients suffering daily with current therapies that are not efficacious in treating the root cause of MI and stroke, atherosclerosis, by reducing inflammation or increasing reverse cholesterol transport (RCT). Atherosclerosis is a vascular inflammatory disease characterized by the formation of fatty plaques which can rupture, leading vessel occlusion. Lymphangiogenesis is the process whereby lymphatic vessels are generated, but its role in atherogenesis is currently under‐characterized and controversial. It has been suggested that lymphangiogenesis is either protective by promoting plaque regression through RCT or it is deleterious by promoting plaque instability through white blood cell trafficking. Our lab has previously reported that Interleukin‐19 (IL‐19), an anti‐inflammatory cytokine, uniquely attenuates atherosclerotic plaque progression while also being pro‐angiogenic. This drives our hypothesis that one mechanism whereby IL‐19 is atheroprotective is by driving lymphangiogenesis, allowing attenuation of plaque progression.Methods & ResultsSeveral approaches were performed to test the hypothesis that IL‐19 is lymphangiogenic. IL‐19 can significantly induce primary human lymphatic endothelial cell (LEC) proliferation, migration, and tube formation, all angiogenic assays, to the same degree as VEGF‐C, a known mediator of lymphangiogenesis. RNA sequencing (RNAseq) analysis indicates that IL‐19 induces chemokine expression as well as an angiogenic transcriptional program in LECs. Notably it induces expression of Prox1, a master transcription factor of lymphangiogenesis, 6.2‐fold. The increases of chemokine and transcription factor expression from RNAseq were verified through qPCR analysis. To further analyze the translational effects of IL‐19, we utilized an LDL Receptor Knockout (LDLR‐/‐) mouse model fed a high fat diet for 16‐weeks with daily injections of either recombinant mouse IL‐19 or saline. Preliminary results suggest an increase in lymphatic vessel presence associated with the known decrease IL‐19 causes in atherosclerotic plaque progression.Conclusion & SummaryThese data suggest that IL‐19 increases lymphatic vessel formation, potentially allowing RCT to take place to decrease plaque burden in atherosclerosis. Further studies will confirm IL‐19’s mechanistic pathway in LECs. We will also determine RCT and inflammatory cell trafficking by intravital microscopy. These results have the potential to shift our understanding toward a positive view of the association between the lymphatic system and atherosclerosis.
Minocycline Blunts Isoproterenol‐Induced Cardiac Hypertrophy
Authors: Russell Jacob , Bailey Chastidy , Brown Scott , DeMarco Vincent G. , Mummidi Srinivas , Bender Shawn B. , Chandrasekar Bysani 
Published: May 2022

IntroductionExcess and chronic activation of the sympathetic nervous system often results in progressive adverse remodeling and heart failure development. Here we investigated whether chronic administration of isoproterenol (Iso), a β‐AR agonist, would promote hypertrophy, fibrosis, and dysfunction, and whether these maladaptive changes are prevented by minocycline, an FDA‐approved antibiotic, that has been shown to have cardioprotective effects outside of its antibacterial properties.MethodsWild‐type male C57BL/6J mice were subcutaneously implanted with osmotic minipumps, delivering either 30 mg/kg body weight per day of isoproterenol, or vehicle control. Following pump implantation, mice received daily intraperitoneal (I.P.) injections of either sterile saline or 50 mg/kg of minocycline. After 21 days, cardiac function was assessed by echocardiography, and hearts were collected for further analysis. A transverse slice of the ventricles was fixed for histology, and a portion of the left ventricle was used for RNA isolation, and mRNA and miRNA sequencing.ResultsEchocardiography revealed that isoproterenol increased heart rate, heart weight (hypertrophy), ejection fraction and fractional shortening, without leading to significant changes in fibrosis. Minocycline prevented the hypertrophic remodeling and changes in systolic function without affecting heart rate. While mRNA sequencing revealed that isoproterenol enriched gene networks associated with changes in inflammation, the miRNA sequencing showed that minocycline enhanced miR‐26b expression, which is reported to suppress GATA4‐mediated hypertrophy.ConclusionMinocycline mitigates isoproterenol‐induced cardiac hypertrophy while preserving contractility, as well as induces changes in RNA and miRNA pathways involved with inflammation and cardiac remodeling
Affinity Maturation of an Ovarian Cancer‐Targeting Peptide
Authors: Asar Mallika C. , Deutscher Susan L. , Soendergaard Mette 
Published: May 2022

Ovarian cancer is among the leading causes of cancer deaths in women. The low survival rate is in part due to inadequate means of detection, which emphasizes the need to develop effective screening techniques. Peptide J18 was previously discovered using phage display technology and was found to specifically target and image human ovarian tumors in a SKOV‐3 xenografted mouse model.Here, affinity maturation of J18 was carried out to improve its binding affinity by conducting an alanine scan of the peptide. A cell‐based modified ELISA using SKOV‐3 cells was carried out and dose‐response curves were modelled using non‐linear regression. The results showed that substitution of serine‐5 (S5) and aspartic acid‐6 (D6) with alanine decreased the EC50 value of the original peptide J18 from 45.02 µM (39.15‐52.94 95% confidence interval; CI) to 19.54 (9.70‐38‐51 95% CI) and 6.92 (5.2‐9.80 95% CI), respectively. Next, a double alanine substitution of S5 and D6 (J18‐S5A‐D6A) was investigated to further increase the binding affinity. The EC50 value was determined to be 4.22 (2.16‐16.14 95% CI), thus improving the binding affinity approximately 10‐fold.To ensure that J18‐S5A‐D6A maintained specificity for ovarian cancer cells, the peptide was evaluated against ovarian cancer SKOV‐3, pancreatic cancer Mia‐Paca‐2, prostate cancer LNCaP, and embryonic kidney HEK 293 cells using a modified ELISA and fluorescent microscopy. Both assays showed significantly increased binding of J18‐S5A‐D6A to SKOV‐3 cells compared to the other cell lines. Based on these results, peptide J18‐S5A‐D6A may be used as an improved imaging agent of ovarian cancer.
Lipocalin‐2 Stimulates Proliferation and a Profibrotic Cholangiocyte Phenotype during Cholestasis
Authors: O'Brien April , Stiles Danaleigh , White Tori , Lopez Matthew , Little Ashleigh , Childs Jonathan , Douthitt Ashley , Phan Thien , Medford Abigail , Chen Lixian , Francis Heather , Alpini Gianfranco , Chakraborty Sanjukta , Glaser Shannon 
Published: May 2022

Lipocalin‐2 (LCN2), also known as neutrophil gelatinase‐associated lipocalin (NGAL), is a secreted glycoprotein that has been shown to regulate cell proliferation, innate immunity, metabolism, and tumor metastasis in several cell types. LCN2 expression is elevated in hepatic tissues in experimental liver injury models and patients with alcoholic hepatitis. LCN2 has been shown to stimulate proliferation and metastasis of cholangiocarcinoma. However, the role of cholangiocyte LCN2 and LCN2 receptors (NGALR/24p3‐R) during cholestasis and primary sclerosing cholangitis (PSC) has not been evaluated. Therefore, we aimed to assess the role of LCN2 and its receptor, 24p3‐R, in acute and chronic cholestatic models.MethodsLCN2 and 24p3‐R were evaluated in liver sections, total liver, and isolated cholangiocytes from wild‐type (WT; C57) normal and BDL (7 days) mice and WT (FVB/NJ) and Mdr2‐/‐ mice (model of primary sclerosing cholangitis, PSC) by immunohistochemistry and qPCR, respectively. LCN2 levels were evaluated by ELISA in cholangiocyte supernatants (6 hrs) collected from animal models and serum from BDL and Mdr2‐/‐ and WT controls and late‐stage PSC patients and healthy human controls. Serum bile acid (BA) levels were measured using a commercial BA assay kit in BDL, Mdr2‐/‐, and control mice. In vitro, both human (H69) and mouse pooled (MPC) cholangiocytes were treated with either 50 ng/ml LCN2 or 100 mM taurocholic acid (TCA) for 24 hrs after which the expression of proliferation, fibrosis, LCN2, and 24p3‐R genes was determined via qPCR.ResultsThere were significant increases in LCN2 and 24p3‐R immunostaining in BDL mice compared to controls. By qPCR significant increases were observed for both LCN2 and 24p3‐R in total liver and isolated cholangiocytes from BDL and Mdr2‐/‐ mice compared to WT. Increased levels of LCN2 were found in serum from PSC patients and serum and supernatants from isolated cholangiocytes from BDL and Mdr2‐/‐ mice compared to control. In vitro, MPC cells treated with LCN2 had increases in fibrotic marker Col1a1, proliferation marker PCNA, and inflammatory markers IL‐1b and IL‐6. Also, significant increases were seen in fibrotic markers (aSMA, FN‐1, and TGF‐b1) and LCN2 receptor 24p3‐R in H69 cells treated with LCN2 compared to untreated cells. As expected, increased serum BA levels were observed in both BDL and Mdr2‐/‐ mice compared to controls. H69 cells treated with TCA also had significant increases in FN‐1, TGF‐b1, and the LCN2 receptor 24p3‐R, indicating that BA may regulate 24p3‐R expression.ConclusionLCN2 and 24p3‐R expression is elevated in cholestatic models and stimulates a profibrotic cholangiocyte phenotype. These findings suggest that LCN2 may play a crucial role in the pathogenesis of cholestatic liver injury.
VCAM1 regulates Fra1/JunB mediated IL‐8 expression and pathological retinal neovascularization in a murine model of oxygen‐induced retinopathy
Authors: Kaur Geetika , Bisen Shivantika , Sharma Deepti , Singh Nikhlesh K. 
Published: May 2022

PurposePathological retinal neovascularization is a clinical manifestation of various proliferative retinopathies, including retinopathy of prematurity (ROP), diabetic retinopathy, and the wet form of macular degeneration. Over the last decade, numerous studies have reported an increased infiltration of hematopoietic stem cells (HSCs), macrophage/microglia in patients with intraocular angiogenic diseases.MethodsUsing a murine model of oxygen‐induced retinopathy (OIR) and human retinal microvascular endothelial cells (HRMVECs), we are trying to understand the significance of IL‐33/Hypoxic induced VCAM1 expression in retinal neovascularization.ResultsHere, using a murine model of oxygen‐induced retinopathy, we found that NFkB and VCAM1 levels were highly up regulated in retina when new vessels were developing in the retina. Genetic deletion of IL‐33 resulted in reduced NFkB activation and VCAM1 levels in hypoxic retina, resulting in reduced neovascularization. These findings suggest a role for IL‐33‐NFkB‐VCAM1 signaling in pathological neovascularization. Our results demonstrate that IL‐33 induce tube formation in human retinal microvascular endothelial cells and depletion of VCAM1 levels significantly blocked it. Protein profiler analysis showed that IL‐33 regulates VCAM1 dependent IL‐8 expression in HRMVECs. In order, to understand how IL‐33 induced VCAM1 regulates the expression of IL‐8, we did sequence analysis of IL‐8 promoter and observed that IL‐8 promoter has AP1 binding site. We observed that IL‐33 induced the expression of Fra1 and JunB significantly in HRMVECs. Cloning and site directed mutagenesis of IL‐8 promoter revealed that the AP1 binds to IL‐8 promoter in response to IL‐33 and regulate the expression of IL‐8 in HRMVECs. Consequently, down regulation of VCAM1 levels significantly reduced IL‐33 induced Fra1 and JunB expression as well as IL‐8 expression.ConclusionsThese results strongly suggest that IL‐33/ST2L‐NFkB‐VCAM1‐AP1 regulates IL‐8 expression, playing a key role in retinal neovascularization and thus represents a new pharmacological target for the treatment of diseases where excessive neovascularization is the underlying pathology.
R‐(‐)2,5‐dimethoxy‐4‐iodoamphetamine (DOI) Blunts the Discriminative Stimulus Properties of Oxycodone
Authors: Salinsky Leah M. , Fox Robert , Anastasio Noelle , Cunningham Kathryn A. 
Published: May 2022

AimsA record ~100,000 fatalities due to drug overdoses was reported in the U.S. for the twelve months preceding April 2021. Opioids account for most deaths, and recurrent opioid use may lead to the development of opioid use disorder (OUD), a debilitating chronic disorder. Current medication‐assisted OUD treatment (e.g., buprenorphine) mitigates withdrawal, reduces mortality, and reduces intake of abused opioids. Still, the opioid epidemic crystalized the need to identify novel non‐opioid therapeutics to improve the odds of successful OUD recovery. In the last decade, there has been a resurgence of interest in serotonergic psychedelics for the treatment of psychiatric disorders, including substance use disorders. Psychedelic compounds act through agonist actions at the serotonin (5‐HT) 5‐HT2A receptor (5‐HT2AR) and there is some literature to suggest functional interactions between 5‐HT2AR and opioids. We tested the hypothesis that the psychedelic 5‐HT2AR agonist R‐(‐)2,5‐dimethoxy‐4‐iodoamphetamine (DOI) will disrupt the discriminative stimulus properties of the commonly misused prescription opioid oxycodone. The drug discrimination paradigm is a rigorous laboratory procedure for modeling the interoceptive effects of drugs and is widely recognized as useful in establishing ligand potency and effectiveness in vivo, as well as interrogating abuse liability.MethodsMale, Sprague‐Dawley rats (n = 13) were trained to discriminate an injection (s.c.) of oxycodone (1 mg/kg) from saline (1 ml/kg) in a two‐lever, water‐reinforced paradigm. Pharmacological testing began upon establishment of the stable oxycodone vs. saline discrimination. Substitution tests with DOI (0‐0.2 mg/kg; s.c.) and combination tests with DOI plus oxycodone were conducted interspersed with maintenance sessions. Percent oxycodone‐lever responding, response rates and latency to the first response were analyzed with an experimentwise error rate of α=0.05.ResultsAll rats acquired the oxycodone vs. saline discrimination to stability. Substitution tests indicated that all DOI doses resulted in percent oxycodone‐lever responding significantly different from the training dose of oxycodone (p< 0.05). Pretreatment with DOI (0.1, 0.2 mg/kg) significantly reduced the percent oxycodone‐lever responding (0.5 mg/kg), relative to pretreatment with vehicle (p < 0.05). The selective 5‐HT2AR antagonist M100907 prevented DOI‐induced suppression of oxycodone‐lever responding (p < 0.05).ConclusionsThe current studies reveal that the psychedelic DOI does not substitute for oxycodone, but does suppress the stimulus effects of oxycodone, an effect which is reversed by the 5‐HT2AR antagonist M100907. Taken together, these studies provide support for expanded interrogation of the efficacy of 5‐HT2AR agonists in counteracting the behavioral effects of opioids.