bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mobile Gyro Control for Intuitive Manipulation of Virtual Anatomy Specimens
Authors: Lin Tai‐Jiun Terry , Zhang Steve , Jeon Sean , Meleveetil Vedanth , Krebs Claudia 
Published: May 2022

The VanVR APP (Vancouver Virtual Reality Anatomy and Pathology Project) is an anatomy educational tool developed as a response to the Covid‐19 pandemic. The web app provided 3D models of scanned prosections to accommodate students with their virtual anatomy lab courses at UBC. The web interface means that interactions with the 3D specimens are controlled by mouse movements, which can be less intuitive and confusing. We have developed a technology that allows students to manipulate and turn the 3D specimens with 6 degrees of freedom using their mobile phones as a controller.The VanVR APP was created using multiple development tools such as Unity Engine for the overall environment setup and UI implementation, C# scripts for the interactive functionalities, 3D photogrammetry for specimen visualization, MongoDB for database, render Unity built WebGL and deployed the web app on Azure app service. To improve the interactivity and increase the intuitive design of the application, a new functionality of mobile gyro control was implemented using Photon to connect with the 3D specimens in WebGL using generated pairing code to match the web app and the mobile device in a virtual room. The orientation of the phone is transmitted to a spherical model in WebGL, which accordingly transforms the coordinates of any chosen specimen displayed in front of the user. This means that simply by tilting a phone like a gyro controller, users will be able to intuitively mirror how the specimen rotates in the virtual lab with significantly more freedom than the traditional mouse movement control. Along with the functionality of zooming in/out and comparing different models side by side, the app simulates the feeling of handling a specimen physically. In addition, learners can view the documented annotations tailored to each specimen illustrating details of various regions on the model.The addition of the mobile gyro control gives a more natural way to examine the specimen. This new feature highlights our student‐centred approach with an emphasis on intuitive and accessible design approaches.The acceleration in the use of virtual technologies during the pandemic has increased the use of related tools and their acceptance in anatomy education. The further development of technologies to improve the student learning experience in AR/VR will certainly broaden accessibility. Some future implementation includes haptic touch feedback on mobile phones which better simulates physical labs, and a mobile AR function that extends the current gyro controller feature, deploying AR rendered specimens assigned from the WebGL end on multiple mobile devices, which can be utilized in a classroom setting.
Ethanolic Extract of Calotropis procera Exhibits Antitumor Effects on Human Breast and Colon Cancer Cells via Cell Cycle Arrest
Authors: Abu‐Gharbieh Eman , Bajbouj Khuloud , Shehab Naglaa G. , Elayoty Salma. M. , Abdel‐Rahman Wael M. , Bustanji Yasser , Sinoj Jithna , El‐Huneidi Waseem 
Published: May 2022

Calotropis procera is an evergreen, perennial shrub of the family Apocynaceae has been used to treat rheumatism, filariasis, and skin disorders, and its leaf has been used to treat jaundice. The flowers’ extract has been used to treat spleen, liver, and stomach problems. Members of the same plant family are reported to exhibit anticancer potential.AimTo assess the flavonoid and phenolic constituents of the plant extract, evaluate the potential antitumor activity of C. procera ethanolic extract and investigate the mechanistic pathway using two different cancer cell lines, namely human breast adenocarcinoma (MCF‐7), human colorectal cancer (HCT‐116) and the cytotoxic effect on a normal human dermal fibroblast (aHDF).MethodsTotal flavonoid and phenolic contents were investigated spectrophotometrically and by HPLC analysis. Cell viability assay, Annexin‐V/PI apoptosis assay, cell cycle analysis and Western blotting were used to evaluate the effect of C. proceraon cytotoxicity, apoptosis, cell cycle arrest and proteins expression profile of the related altered pathways.ResultsCoumaric acid was found to be the principal constituent in the ethanolic extract. Quercetin and hisperdin were the majour flavonoids. The extract demonstrated potential and effective cytotoxic activity against the cancer cell lines, MCF‐7 and HCT‐116, with a minor toxic effect on normal cells. The extract significantly induced apoptosis and cell cycle arrest at the G2/M phase in both cancer cells. Western blot analysis revealed a significant reduction in G2/M regulators, cdk1 and cyclin B1, and the proliferation marker, survivin, protein levels in the treated cells.ConclusionThis study highlights evidence of the novel antitumor effect of C. procera in breast and colon cancers by altering the progression of the cell cycle.
Resveratrol Reduces Arterial Stiffness and Improves Functional Capacity in Patients with COPD
Authors: Cobb Kolton , Payne Christian , Lavender Rebekah , Simovic Tijana , Harris Ryan A. , Pollock Jennifer , Baban Babak , Mannino David , Nana‐Sinkam Patrick , Rodriguez Miguelez Paula 
Published: May 2022

IntroductionChronic Obstructive Pulmonary Disease (COPD) is the 3rd leading cause of death worldwide and is strongly associated with the development of cardiovascular disease (CVD). Large artery stiffening is recognized as a major CVD risk factor that contributes to diminished myocardial blood supply and reduced functional capacity, a marker of disease severity in COPD. Elevated oxidative stress plays a pivotal role in the etiology of both COPD and CVD. Apart from the body’s endogenous ability to counteract oxidative stress, the consumption of exogenous antioxidants also enhances the capacity to scavenge free radicals and reduce arterial stiffness. Whether or not antioxidants can reduce arterial stiffness and improve functional capacity in patients with COPD, however, has yet to be elucidated.PurposeThis study sought to test the hypothesis that oral resveratrol, a powerful antioxidant, will reduce arterial stiffness and improve functional capacity in patients with COPD.MethodsTen patients with COPD (GOLD Stage II‐IV) participated in this study and completed 6 weeks of oral resveratrol (500 mg/daily) or placebo (PLC) treatment. Arterial stiffness was evaluated via tonometry (SphygmoCor) before and after RES or PLC to assess augmentation index (AIx), AIx normalized to a heart rate of 75 beats per minute (AIx75), and diastolic pressure time index (DPTI). Functional capacity was measured via six‐minute walk test to evaluate distance walked.ResultsA single dose of RES significantly (p<0.01) improved markers of arterial stiffness (AIx75: ∆RES=‐8±2% vs. ∆PLC=‐4±2%) while increasing myocardial perfusion (DPTI: ∆RES=158±79 mmHg vs. ∆PLC=‐23±18 mmHg p=0.04). No changes (p=0.171) in functional capacity were observed after a single dose of RES or PLC. However, six weeks of RES significantly (p=0.01) reduced AIx75 (∆RES=‐9±3% vs. ∆PLC=‐3±2%), while it improved myocardial perfusion (DPTI: ∆RES=166±85 mmHg vs. ∆PLC=‐36±18 mmHg; p=0.04) and distance walked (∆RES=25±13m vs. ∆PLC=‐2±4m; p=0.04).ConclusionFor the first time, we have identified that the antioxidant resveratrol reduces markers of arterial stiffness and improves myocardial perfusion in patients with COPD. In addition, six weeks of daily intake of this natural antioxidant also improved functional capacity in this population. Future studies are warranted to investigate the long terms effects of resveratrol in cardiovascular health in patients with COPD.
Systemic proteomic and metabolomic signature of the SGLT2 inhibitor dapagliflozin
Authors: Billing Anja , Kim Young‐Chul , Vallon Volker , Rinschen Markus 
Published: May 2022

BackgroundDapagliflozin is a member of a new class of medication called gliflozins indicated for the treatment of diabetes. Gliflozins lower blood glucose levels by inhibition of the sodium glucose co‐transporter 2 (SGLT2), which is predominantly expressed in the S1 segment of renal proximal tubules and responsible for >90% of renal glucose reabsorption. Gliflozins also protect the kidney and heart, which cannot be solely explained by blood glucose lowering. To gain further mechanistic insights, we determined the metabolomic and proteomic signature of dapagliflozin in a murine model of diabetes.MethodsMale C57BL/6 non‐diabetic wildtype and littermate Ins2 (Akita) diabetic mice, both fed a Western diet, were given dapagliflozin (10 mg/kg diet) or vehicle for one week (4 groups, N=8/group). Blood and urine glucose levels confirmed the expected treatment response. We analyzed metabolome, proteome and phosphoproteome for a range of tissues (kidney, liver, heart, smooth muscle, white adipose tissue) and body fluids (serum, urine, erythrocytes), and extended the analysis to the gut metaproteome. For untargeted metabolomics, analytes were measured by HILIC‐MS/MS and RP‐MS/MS. For proteomics and phosphoproteomics, TMT labelled (TMT16plex) peptides were separated by high pH reverse phase fractionation and measured by LC‐MS/MS including FAIMS (Thermo Scientific Orbitrap Exploris 480 mass spectrometer).ResultsMulti‐layered omics data were obtained comprising 11341 proteins, 14762 phosphorylation sites, and 217 metabolites. In general, dapagliflozin‐induced changes were more distinct in Western diet‐fed non‐diabetic than diabetic mice. Metabolomics revealed effects of dapagliflozin on amino acid, carbohydrate and lipid metabolism with most changes observed in urine, serum and liver. In comparison, dapagliflozin‐induced perturbations on proteome and phosphoproteome level were observed predominantly in kidney and affected ATP/lipid/carbohydrate/nitrogen metabolism, mitochondria, mitophagy, methyl transferases, methyl CpG‐binding, kinases, deacetylases, endosomes, cell junctions and gene expression. Interestingly, also the gut metaproteome was affected by dapagliflozin with significant changes in Bacteroides and Bifidobacterium species.ConclusionThe SGLT2 inhibitor dapagliflozin induces distinct effects on kidney proteomics, associated with robust metabolomic signatures in plasma, urine and liver, as well as effects on the gut metaproteome. These effects occurred independent of blood glucose levels and support a concept of pleiotropic effects of SGLT2 inhibition that may originate in the kidney but induce wide‐ranged extrarenal metabolic effects and consequences.
Development of an Automated, High‐Throughput Assay to Detect Angiotensin AT2‐Receptor Agonistic Activity by Nitric Oxide Measurement In Vitro
Authors: Silva Igor M. S. , Kjærgaard Kenneth , Mortensen Christina , Stage Tore B. , Steckelings Ulrike M. 
Published: May 2022

BackgroundThe angiotensin AT2‐receptor (AT2R) is a key component within the protective arm of the renin‐angiotensin system inducing vasodilation by nitric oxide (NO) production. Currently, no high‐throughput assay is available to identify AT2R agonists in vitro, which could explain the low number of AT2R selective ligands in drug development programs.ObjectiveTo design and validate a high‐throughput method for detection of AT2R activation in vitro based on NO release.MethodsHuman aortic endothelial cells (HAEC) were seeded into 96‐well plates at a density of 5000 cells per well. 48 hours after seeding, cells were loaded for 30 minutes with the NO probe, DAF‐FM diacetate (5µM). Cells were stimulated for 15 minutes with the AT2R‐agonist C21 (1µM) or with classical NO‐releasing agents as positive controls: bradykinin (BK, 0.1µM), acetylcholine (ACh, 1µM) or Ang‐(1‐7) (MasR agonist, 0.1µM). Cells were fixed and fluorescence signals captured by fluorescence microscopy with automated, built‐in image analysis (ImageXpress Pico, Molecular Devices, USA). AT2R‐transfected (AT2R‐CHO) or non‐transfected (NT‐CHO) Chinese hamster ovary cells were cultured and loaded with DAF‐FM diacetate according to the same protocol as HAEC. Concentration‐response curves were obtained for C21, BK, Ang II and Ang‐(1‐5) (0.1nM to 10µM each) using the automated protocol for ImageXpress Pico described above on AT2R‐CHO. To evaluate whether effects of the compounds in AT2R‐CHO were AT2R specific, the response to these compounds at the highest concentration (10µM) was also tested in NT‐CHO.ResultsUsing primary HAEC we could show that the automated, high‐throughput workflow was able to detect NO release by AT2R agonists as efficiently as by classical NO‐releasing agents [C21: 16.16 ± 6.84%; Ang‐(1‐7): 25.45 ± 3.37%, BK: 19.30 ± 2.92%; ACh: 24.01 ± 4.24%; one‐way ANOVA followed by Dunnett’s multiple comparison test, p≤0.05 vs control, ≥3 independent experiments for all compounds). In AT2R‐CHO, we detected that both C21 and Ang II (AT2R agonists) increased NO release in a concentration‐dependent manner, while BK had no effect (AT2R‐CHO lacks BK receptors) at any tested concentration. Surprisingly Ang‐(1‐5), a degradation product of Ang II thought to be inactive, induced NO release in a concentration‐dependent manner in AT2R‐CHO. In NT‐CHO, none of the compounds stimulated NO release, indicating that the effects by C21, Ang II and Ang‐(1‐5) in AT2R‐CHO were AT2R specific.ConclusionMeasurement of NO release from AT2R‐CHO or HAEC by DAF‐FM diacetate in an automated way is suitable as high‐throughput assay for the identification of AT2R‐agonistic compounds in vitro. Application of the assay revealed that Ang‐(1‐5), which has been regarded as an inactive metabolite of Ang II, has AT2R agonistic properties.
Chronic Intermittent Hypoxia Alters Chloride Gradients in Median Preoptic Nucleus (MnPO) Neurons of Rats: Comparing ClopHensorN and Perforated Patch Recording
Authors: Farmer George E. , Little Joel T. , Cunningham J. T. 
Published: May 2022

Chronic intermittent hypoxia (CIH) is an animal model that simulates the hypoxemia associated with sleep apnea. Male rats exposed to CIH exhibit increased sympathetic tone and reactivity and persistent elevations in blood pressure. In MnPO neurons, CIH induces reductions in GABAa inhibition that are, in some neurons, exacerbated to the point of GABAa mediated excitation. The genetically encoded ratiometric Cl‐ sensor, ClopHensorN, was used to monitor the Cl‐ flux of PVN projecting MnPO neurons in normoxic (Norm) and CIH treated rats in response to GABAa receptor activation. In some experiments, ClopHensorN imaging was combined with perforated patch recordings.Using isoflurane (2‐3%) anesthesia, male Sprague‐Dawley rats (250‐350g) received bilateral infusions (0.4 µL) of a retrograde AAV9‐Cre in the PVN and a DIO‐ClopHensorN in the MnPO. After recovery, rats were subjected to 7 days of CIH (0800‐1600 hr) or normoxia. CIH consisted of 6 min cycles (3 min 21% O2, 3 min 10% O2) repeated 10x/hr for 8 hours (during the normal inactive/sleep phase) on 7 consecutive days. For recordings, rats were anesthetized with isoflurane (2‐3%) and coronal slices (300 µm) containing the MnPO were cut using standard in vitro slice procedures. All cells were tested in standard artificial cerebrospinal fluid for their responses to 10 s focal application of muscimol (100 uM). ClopHensorN images were captured every 3 sec. Cl‐ flux was determined from the ratio of GFP (ex 488 nm, em 500‐550 nm) to tdTomato (ex 594 nm, em 650‐700 nm) fluorescence. For some cells, voltage‐clamp recordings using gramicidin (100 ug/ml in 140 mM K‐gluconate) perforated patch were also collected (in labeled and unlabeled MnPO neurons).MnPO neurons from CIH treated rats (n = 632), 20.1% showed a decrease in the ratio of GFP to tdTomato fluorescence (n = 127) while 0.3% showed increases in this ratio (n = 2) indicative of Cl‐ efflux. In MnPO neurons from Norm rats (n = 482), 41.9% showed a muscimol dependent decrease in the ratiometric Cl‐ signal (n = 202) with 0 cells showing increases. Muscimol dependent decreases in the Cl‐ signal were reduced in the CIH treated rats suggesting reduced GABAa inhibition. During perforated patch recordings, MnPO neurons from Norm rats exhibited mIPSCs and outward currents in response to muscimol. MnPO neurons from CIH treated rats show variable mIPSC frequencies and heterogeneous responses to muscimol.The results demonstrate that CIH alters the Cl‐ flux of PVN projecting MnPO neurons in response to GABAa stimulation. These changes may contribute to the increased sympathetic tone and reactivity, as well as the persistent hypertension associated with CIH.
Gut microbiota depletion with antibiotics enhances fructose induced salt‐sensitive hypertension in normal rats
Authors: Haque Mohammed Z. , Rey Federico , Ortiz Pablo A. 
Published: May 2022

Introduction‐We reported that 20% fructose in drinking water induced salt‐sensitive hypertension in normal rats. High fructose (HF) or high salt (HS) intake reportedly alter the gut microbiota, which influences a variety of microbial‐regulated bioactive metabolites that can affect blood pressure and renal function. The contribution of the gut microbiota to fructose induced salt‐sensitive hypertension or renal function has not been studied. Aim‐ We hypothesized that gut microbiota depletion with antibiotics (ABX) would change fructose induced salt‐sensitive hypertension, renal excretory function and plasma levels of bacterial metabolites (lactate, β‐hydroxy‐butyrate).Methods‐Sprague Dawley rats were treated with a cocktail of antibiotics (ABX group, in g/L 1.0 ampicillin, 1.0 neomycin sulphate, 1.0 metronidazole, and 0.5 vancomycin in drinking water) for 4 weeks followed by a 0.33 fractional maintenance dose for up to 12 weeks) or control diet (no‐ABX). On week 5, all rats were given 20% fructose (HF) in drinking water and fed a diet with normal salt (NS, 0.4% Na) or high salt content (HS, 4% Na). Rats were divided as: Group 1 (HF + NS, no‐ABX, n= 6), Group 2 (HF + NS + ABX, n=6), Group 3 (HF + HS, no‐ABX, n= 8), Group 4 (HF + HS + ABX, n= 8). We trained rats and measured systolic blood pressure (SBP) for 12 weeks by tail‐cuff and collected feces before and after ABX, to measure gut c‐DNA content. On the 12th week, we collected 24‐ hour urine and plasma before the end of the protocol.Results‐ Antibiotics induced microbiome depletion (ABX) decreased fecal DNA content by 83%. Baseline SBP (in mmHg) or urine albumin excretion was not affected by ABX. Fructose with normal salt diet did not change SBP with (125±4) or without ABX (124±3). HS significantly increased SBP in rats fed fructose without ABX (from 123±3 to 141±4, p<.05) but the increase was greater in ABX rats (from 124±4 to 155±3, p<.05). Average increase on SBP was 26±2 in HF+HS with ABX, but only 15±2 without ABX (p<.001). In HF + HS groups, ABX decreased plasma lactate from 1657±259 to 905±144 µM (p<.01), and β‐hydroxy‐butyrate from 861±144 vs 220±53 µM (p<.001). Non‐fasted plasma insulin was also decreased by ABX in HF+HS rats (0.77±0.13 to 0.36±0.06 ng/mL, p<.03). The increase in body weight was similar in HF+HS rats with or without ABX.Conclusion‐ Chronic antibiotic treatment induced gut microbiota depletion and enhanced fructose induced salt‐sensitive hypertension in normal rats, an effect that was associated with decreased plasma beta‐hydroxy‐butyrate, lactate and insulin. Additional studies should focus on direct renal effects of ABX on salt handling.
Urinary Serpin‐A3 is an Early Predictor of Clinical Response to Therapy in Patients with Proliferative Lupus Nephritis
Authors: Martínez‐Rojas Miguel Angel , Sánchez‐Navarro Andrea , Mejía‐Vilet Juan Manuel , Pérez‐Villalba Rosalba , Uribe Norma , Bobadilla Norma 
Published: May 2022

BackgroundSerpin‐A3 is a regulator of different inflammatory processes, however its role in the kidney is unknown. We previously reported that urinary Serpin‐A3 excretion (uSerpA3) is significantly elevated in patients with active lupus nephritis (LN) and rats with Chronic Kidney Disease (CKD).ObjectiveDetermine the association between uSerpA3 temporal course during the 1st‐year of treatment and response to therapy in patients with proliferative LN.HypothesisuSerpA3 will correlate with kidney inflammation and will predict poor response to therapy.MethodsThis is an observational longitudinal study including sixty Mexican adults with proliferative LN followed during the 1st‐year after LN‐flare. uSerpA3 at flare and after 3, 6, and 12 months was evaluated semiquantitatively through Western blot; histological evaluation was performed by kidney biopsy. Outcome: Response to therapy 1‐year after the LN‐flare. Relation between uSerpA3 and histopathologic characteristics at flare was evaluated cross‐sectionally; the temporal association between uSerpA3 and response to therapy was analyzed with linear mixed models; prognostic performance for clinical response was evaluated with ROC analysis.ResultsAmong the 60 patients studied, 21 (35%) were class III and 39 (65%) class IV. The class IV LN had higher serum creatinine, lower C3 levels, and higher histological activity and chronicity indexes. The uSerpA3 was higher in class IV than in class III LN (6.98 vs. 2.89 DPI/mg‐creatinine, p=0.01). Furthermore, uSerpA3 had a mild correlation with the histological activity index (r=0.29, p=0.02). There was a significant association between the temporal course of uSerpA3 and the response to therapy. Responders showed a significant drop in uSerpA3 at 6th‐month compared to LN‐flare (p<0.001), while non‐responders persisted with elevated uSerpA3. Moreover, uSerpA3 was significantly lower at flare in responders compared to non‐responders (2.69 vs 6.98 DPI/mg‐creatinine, p<0.05). Furthermore, uSerpA3 was able to identify non‐responders since the 3rd‐month after LN‐flare, (AUC=0.77).ConclusionsuSerpA3 is an early indicator of kidney inflammation and a potential predictor of long‐term response to therapy in patients with proliferative LN.
Renal inflammatory signaling alters cardiomyocyte kinetics and lead to HFpEF in CKD: A mechanistic proof of concept study
Authors: Chade Alejandro R. , Mohamed Tamer M. , Eirin Alfonso 
Published: May 2022

BackgroundChronic kidney disease (CKD) carries a higher cardiovascular mortality and is a prominent risk factor for development of HFpEF. However, the pathophysiology of HFpEF in CKD has not been established. We have recently shown that experimental CKD‐HFpEF is associated with increased renal release of inflammatory cytokines, particularly tumor necrosis factor (TNF)‐α and interleukin (IL)‐6, to the systemic circulation. Using a unique physiological biomimetic heart culture system, we test whether inflammatory cytokines of kidney origin contribute to the development of HFpEF in CKD.MethodsPigs (4 each) were assigned to sham or CKD (bilateral renovascular disease and dyslipidemia) groups and followed for 14 weeks. Circulating levels of TNF‐α and IL‐6 were quantified (ELISA). In a biomimetic heart culture system, pig heart slices were exposed to plasma from normal or CKD pigs, or from normal pigs enriched with TNF‐α and IL‐6 (matching CKD concentration) with or without cytokine neutralizing antibodies (NA). Contractility and relaxation kinetics in vitro were determined in a dose/time dependent manner.ResultsCKD‐HFpEF pigs exhibited elevated systemic levels of TNF‐α and IL‐6 (Figure 1A), associated with renal dysfunction, cardiac hypertrophy and fibrosis, and diastolic dysfunction. Plasma was collected from all animals. Then, pig heart slices were exposed to CKD‐HFpEF, normal, or normal plasma enriched with cytokines, showing impaired contractility and speed of relaxation that improved after TNF‐α and IL‐6 neutralization (Figure 1B).ConclusionsOur observations implicate circulating inflammatory mediators released by dysfunctional kidneys in imposing damage to the remote myocardium, supporting a role of inflammatory cytokines of renal origin in renal‐cardio pathophysiology in CKD‐HFpEF.