bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural Heteropolysaccharide Adhesion to the Corneal Glycocalyx
Authors: Liu Betty S. , Liao Matthew , Wagner Willi , Khalil Hassan , Chen Zi , Ackermann Maximilian , Mentzer Steven J. 
Published: May 2022

The cornea presents unique challenges to wound care. The cornea is constantly exposed to bacterial colonization, has ongoing risk of wound‐related astigmatism, and is cumbersome for traditional wound dressings. A potential solution to these challenges is provided by the corneal glycocalyx.Recent high pressure rapid freezing fixation techniques suggest that the glycocalyx is much thicker than previously believed. In the case of the cornea, the glycocalyx may provide an opportunity for wound care anchored to the transmembrane carbohydrate surface layer.To probe the surface glycocalyx we used a plant‐derived structural heteropolysaccharide called pectin. Pectin has been shown to mirror the structure of the glycocalyceal surface layer of visceral organs. To test the adhesivity of pectin to the corneal glycocalyx, high methoxyl citrus pectin was molded into tape form. Bovine globes were obtained from a local abattoir and used same day. Experiments were performed with custom fixtures using a materials analyzer (TA‐XT plus, Stable Micro Systems). To assess pectin adhesion to cornea, pectin was compressed onto the corneal surface then withdrawn, recording force and distance. A corneal incision was sealed with pectin and subject to increased pressure, until leak. At least three biologic replicates were analyzed, expressed as mean ± one standard deviation. Significance was assessed by Student’s T‐test. Pectin films on cornea were also fixed with formalin and imaged using a scanning electron microscope (Philips XL30 ESEM).Pectin films were flexible, translucent, thin (80um), and did not distort an Amsler grid. Pectin was significantly more adherent (3.2±0.54 N) to the bovine cornea than control polymers nanocellulose fibers (1.2±0.4 N), sodium hyaluronate (1.7±0.7 N), and carboxymethyl cellulose (1.3±0.1)(p<0.05). The work of adhesion, reflecting the area under the adhesion curve, was also significantly greater (p<0.05, asterisk, Figure 1). The cornea incisions sealed with pectin were resistant to pressure fluctuations ranging from ‐51.3±8.9 mmHg to +214.4±68.0 mmHg, both extremes beyond physiologic human intraocular pressures. Sudden or complete sealant failure was not observed. Consistent with these findings, scanning electron microscopy demonstrated a low‐profile tape densely adherent to the bovine cornea (Figure 2).The tensile and peel adhesive strength of the pectin biopolymer suggests a mechanism of polymer‐polymer entanglement with the corneal glycocalyx. As a therapeutic option for corneal wounds, the entangled pectin provides not only a protective barrier, but also an opportunity for pectin‐embedded drug delivery.
HIF1α and NFκB Regulate Mineralocorticoid Receptor Gene Expression in Aging Human Vascular Smooth Muscle Cells
Authors: Ibarrola Jaime , Lu Qing , Zennaro Maria‐Christina , Jaffe Iris Z. 
Published: May 2022

BackgroundMineralocorticoid receptor (MR) activation drives aging‐associated cardiovascular diseases including hypertension, vascular stiffness and heart failure. Vascular MR expression increases with age and drives the aging phenotype but the mechanism of MR gene regulation is unknown. The MR has two mRNA isoforms (1α and 1β) that differ in only the 5’‐untranslated region (exon 1α or 1β) and in the promoter sequence (P1 or P2, respectively). Thus, we investigated the transcriptional mechanisms regulating MR gene expression in aging human smooth muscle cells (SMC).MethodsMR mRNA and protein expression were quantified in primary human aortic SMC (HASMC) and human aortic tissue comparing adult (average age 40s) and aged (average age 70s) male and female donors and comparing adult HASMC or Pac1 (rat SMC line) treated with H2O2 to induce aging. MR promoter activity was tested by luciferase reporter assays. In silico analysis of the MR promoters identified putative transcription factor (TF) binding sites and inhibitor studies and chromatin immunoprecipitations (ChIP) were performed to validate the predicted regulatory mechanisms.ResultsIncreased SMC donor age or H2O2 treatment were associated with significantly increased expression of both of the MR mRNA isoforms, MR protein level, and activity of both MR promoters in HASMCs. The hypoxia‐inducible TF, HIF1α, and the inflammatory TF, NFκB, were predicted in silico to bind to the MR promoters and found to increase significantly with age in HASMCs. HIF1α was induced by H2O2 in HASMCs and this was associated with significant enrichment of HIF1α binding specifically at a site in the 1a exon, near the MR P1 promoter. HIF1α inhibition attenuated expression specifically of the MR‐1α mRNA isoform resulting in decreased MR protein in HASMC. NFκB also increased with age in HASMCs and contributes to MR expression. In response to H2O2,NFκB binding is enriched at to two different binding sites, one each in the P1 and P2 promoters and NFκB inhibition decreased expression of both MR mRNA isoforms (1α and 1β) and MR protein in SMC. Finally, HIF1α and NFκB expression each positively correlated with donor age and with MR expression (p<0.0001) in human aortic tissue from males and females.ConclusionThe MR gene is regulated by the inflammatory transcription factor NFkB and by the hypoxia/oxidative stress‐induced TF HIF1α, to increase MR expression in SMC of the aging human vasculature. This molecular insight may explain enhanced MR activity in cardiovascular aging and could potentially contribute to other cardiovascular conditions driven by oxidative stress and inflammation.
The LP07 and LLC pre‐clinical models of lung cancer present divergent anabolic deficits and expression of pro‐inflammatory effectors of muscle wasting
Authors: Belcher Daniel J. , de la Rosa Guitart Maria , Hain Brian , Kim Hyo‐Gun , Waning David , Barreiro Esther , Nader Gustavo A. 
Published: May 2022

AbstractPre‐clinical models of lung cancer have been instrumental in uncovering the mechanisms of muscle wasting. However, differences in genetic background, immune status, tumor origin, and administration route can introduce confounding variables, limiting the understanding of muscle wasting in lung cancer. In this study, we examined anabolic deficits and the expression of pro‐inflammatory cytokines in the skeletal muscle of LP07 and Lewis lung‐cell carcinoma (LLC) pre‐clinical models of lung cancer. Female mice were injected with either LP07 or LLC tumor cells in their right flank. Tumor growth occurred over a 28–30‐day period and resulted in gastrocnemius muscle depletion with more pronounced wasting in the LLC model (LP07: ‐20.3%, P=0.004; LLC: ‐31.9%, P<0.001). The LP07 mice experienced a loss in ribosomal (r) RNA (‐34.9%, P=0.015), which was likely due to decreased rDNA transcription elongation, evidenced by a lower internal transcribed spacer (ITS) signal (‐47.9%, P=0.017). Reductions in RPS6 (LP07: ‐60.8%, P=0.020; LLC: ‐83.0%, P=0.003) and 4E‐BP1 (LP07: ‐31.4%, P<0.001; LLC: ‐36.5%, P<0.001) phosphorylation was similar in both models but was independent of changes in mTOR phosphorylation which was less pronounced in the LP07 (‐2.8%, P=0.997) compared with the LLC model (‐44.2%, P=0.042). Additionally, IL‐6 and TNF‐α mRNA levels were similarly elevated in the LLC (IL‐6: 750.7%. P<0.001; TNF‐α: 156.5%, P<0.001) and LP07 models (IL‐6: 719.4%, P<0.001; TNF‐α: +100.0%, P=0.28). In summary, while both models underwent muscle depletion, only the LP07 model displayed a reduction in rRNA and rDNA transcription. RPS6 and 4E‐BP1 phosphorylation’s were similar in both models, however, the increase in pro‐inflammatory factors was more pronounced in the LLC model. Our findings reveal a significant divergence in the anabolic deficits and expression of pro‐inflammatory effectors of muscle wasting in the LP07 and LLC pre‐clinical models of lung cancer.
Cytokine and Chemokine Release from Immortalised Human Myotubes in Culture: Effect of Contractions and Comparison with Isolated Intact Mature Adult Murine Muscle Fibres
Authors: Neiland Harvey J. , Staunton Caroline A. , Owen Euan , Kumiscia Jekaterina , Shigdar Shahjahan , Jones Samantha W. , Jackson Malcolm J. , McArdle Anne 
Published: May 2022

Skeletal muscle releases a number of signalling molecules, including immunomodulatory cytokines and chemokines which can act in an autocrine, paracrine and endocrine fashion. For example, neuromuscular integrity is dependent on efficient and appropriate signalling from muscle to nerve and vice versa and when such signalling fails then tissue deterioration occurs. The aim of this study was to characterise the pattern and time course of release of pro‐ and anti‐ inflammatory cytokines and chemokines from cultured immortalised human myotubes at rest and following contractions compared with release from isolated viable intact mouse muscle fibres. Immortalised human myoblasts were fused to mature into myotubes for 7 days (Donna et al, 2003). Alternatively, single muscle fibres were isolated from the flexor digitorum brevis (FDB) muscle of adult C57Bl/6 mice (Pye et al, 2007). Cells and isolated fibres were subjected to electrical field stimulation with trains of biphasic square wave pulses of 2 ms duration for 0.5 s repeated every 5 s at 50 Hz and 30 V per well of fibres for a total stimulation time of 15 min. Media from all samples was analysed for cytokine content at 15 minutes and 24 hours following contractions using a multiplexed cytokine assay (Luminex) and data were standardised to protein content of the well. At 15 minutes following contractions of isolated muscle fibres, a significant increase in a number of cytokines and chemokines were seen in the media, including IL‐5, IL‐6, IL‐10 and G‐CSF although this was not fully replicated in 2D cultured myotubes. At 24 hours following contractions, IL‐6 content was significantly increased in the media of both isolated fibres and 2D cultured myotubes. Thus, data demonstrate some commonality of cytokines/chemokines released but future studies will examine release from more physiological 3D muscle constructs as well as the effects of such release on neuronal cell structure and function.Pye et al, J Physiol. 2007 May 15; 581(Pt 1): 309–318.Donna et al, Mol Cancer Res. 2003 (1) (9) 643‐653.
Sex Differences in the Relationship between Exercise and the Brain
Authors: White Zach , Hall Stephanie 
Published: May 2022

IntroductionExercise is an effective, non‐pharmacological therapy to promote brain health. Sex differences in cognitive changes through the lifespan and in response to exercise training are documented, yet more information is needed to characterize these disparities. We hypothesized that aerobic exercise training would improve memory, coordination, and muscular strength in female rats and have no significant effect in male rats.MethodsThirty‐two Fisher 344 animals (16 males and 16 females) were housed 2 per cage, on a 12:12 light:dark cycle with food and water ad libitum. At two months old, following a 1‐week acclimatization period where the non‐compliant runners were identified, they were randomly assigned to sedentary or exercise training groups. Those in the exercise training group completed a 10 week treadmill training protocol. The protocol was administered during the animal’s dark cycle and began at 15 cm/s for 15 minutes and increased up to 35 cm/s for 60 minutes. Animals in the sedentary group were restricted to cage activity. Memory (Morris water maze), coordination (Rotarod) and muscular strength (grip strength) were assessed following the exercise intervention.ResultsA significant exercise training effect was seen in females but absent in males. Female runners showed greater performance in the Morris water maze (9.2 sec. vs 32.7, p <.05) and in the Rotarod (69 sec. vs 47.5, p <.05). No significant training effect was documented in the male groups. Also, no training effect was documented in grip strength in either males or females.ConclusionIn the present study, memory and coordination improvements as a result of aerobic exercise training were seen in female but not male rats. This work adds to existing literature which investigates sex differences in cognitive health and adaptations to exercise training.
Riding a VeSPha in the classroom: A Versatile Simulation tool for effective didactic illustration of Pharmacodynamic concepts
Authors: Iniguez‐Lluhi Jorge 
Published: May 2022

A solid mastery of Pharmacodynamics concepts is a cornerstone of effective pharmacology education. Understanding the meaning and appropriate use of pharmacodynamics terms such as potency, affinity, intrinsic efficacy, or receptor reserve is extremely valuable but challenging for most trainees. The onus, therefore, is on the educators to devise effective strategies and tools to facilitate the acquisition of this critical competency.When approaching pharmacodynamics, the ability of learners to explore the influence of driving parameters on the magnitude and position of concentration response curves is a powerful way to visualize and make concrete the basic concepts of pharmacodynamics. To this end, I have developed VeSPha, a Versatile Simulation tool for Pharmacodynamics based on a rapid equilibrium generalization of Furchgott’s views on agonism. Through multiple sliders, the user can vary the parameters of the system, such as the concentrations of orthosteric and allosteric ligands, their respective affinities, the allosteric coupling constant, the intrinsic efficacies of each of the receptor species, the total number of receptors and the sensitivity of the system to the stimulus. A graph panel allows the display of the resulting response curves as a function of the concentration of ligand as well as orthosteric and allosteric ligand occupancy curves. Dropdown lines with corresponding EC50 as well as apparent Kd values for each ligand are also displayed. VeSPha also includes bar graphs as well as an intuitive graphic representation of the model displaying the values of the parameters, the distribution of the receptor species at the chosen state, the total stimulus as well as the resulting responses.The tool allows easy exploration of a wide gamut of possibilities that illustrate basic concepts of affinity, intrinsic efficacy, partial agonism, constitutive activity, inverse agonism, competitive antagonism, as well as allosteric modulation of both affinity and efficacy in any combination. The tool is implemented as an easy‐to‐use Excel file. This allows for flexible standalone use and for straightforward embedding into PowerPoint presentations for real‐time illustrations during lectures. The tool also readily generates simulated data including sampling errors for composing realistic, internally consistent assessment items such as quiz, problem set and exam questions.Implementation of this didactic tool is straightforward and effective in the author’s experience across a wide range of learners spanning from undergraduates, Master’s, Ph.D. M.D. Pharm D and DDS students. The tool, including a tutorial video can be made available to the Pharmacology teaching community upon request and an online version is under development.
Vaccine Immunogenicity assay in bovines against fasciola hepatica
Authors: Zambrana Ana I. , Checa Jackeline , Goyeche Antonella , Alonzo Pablo , Silveira Fernando , Cancela Martín , Maggioli Gabriela 
Published: May 2022

Fasciolosis affects cattle health and production around the world, resulting in economic loss for farmers. Current treatment against this parasite in cattle is based in drugs, which do not prevent infection and/or liver damage. In this work, we present the results of immunogenicity assays by ELISA which will lead to vaccine trials against F. hepatica with a Uruguayan and native plant derived adjuvant. Using four different vaccine formulations and a total of 27 cows, my PhD project currently aims to select the best formulation through ELISA studies from blood samples collected in 2021. The novel natural adjuvant production system (ISCOM) shows a promising replacement for current synthetic adjuvants due to its protective properties. Also, the vaccine formulations are produced with recombinant proteins that are present in the parasite (FhLAP1, FhLAP2, FhStf1 and FhStf2).With these results that I will present in my poster, I will further analyze samples through RNA seq. so that I can select the best formulation for a first vaccine trial and fasciola hepatica infection in 2023.
Augmented Urinary Excretion of Angiotensinogen Precedes Albuminuria in Human Young Adults with Overweight and Mice with High Fat Diet‐Induced Type 2 Diabetes
Authors: Gonzalez Alexis A. , Rivera Patricia , Miranda Catalina , Reverte Virginia , Prieto Minolfa C. 
Published: May 2022

Obesity is linked to the development of hypertension and type 2 diabetes (T2D) and both causing chronic kidney disease (CKD). Increased body mass index (BMI) and high fat diet (HFD) contribute to activation systemic renin angiotensin system (RAS). In addition, activation of intrarenal and intratubular RAS has been associated with increased pressure (BP), CKD, and augmentation of urinary excretion of angiotensinogen (uAGT) during T2D. However, whether augmentation of uAGT reflect intrarenal/intratububular RAS activation in subjects with overweight, remains unclear. Here, we tested the hypothesis that increases in uAGT precedes renal dysfunction in mice and human subjects with overweight,MethodsC57BL/6 mice (N = 10) were fed either a normal fat diet (NFD) or HFD (45% Kcal from fat) for 28 weeks. During this period, metabolic phenotype along with urinary albumin, creatinine, reactive oxygen species (ROS) and AGT, were measured every 4 weeks. In addition, for evidence in humans (age 19‐22 years‐old), 11 individuals with overweight and 17 individuals with normal weight, were studied. In these subjects, anthropometric parameters, systolic and diastolic BP, fasting blood glucose (FBG), and urinary mRNA levels of AGT and kidney injury markers IL‐18 and connecting tissue growth factor, were determined. Patients with history of cardiac or cerebrovascular events and endocrine diseases, anti‐hypertensive treatment (ACE inhibitors, ARBs) and/or anti‐diabetic drugs, were excludedResultsIn mice with metabolic alterations overtime, systolic BP significantly increased after 18 weeks in HFD but not NFD mice. Intrarenal ROS increased in HFD mice. These changes paralleled the augmentation uAGT excretion, which occurred in the absence of overt albuminuria. In human subjects with overweight, systolic and diastolic BP, urinary transcripts levels of AGT, renin, and injury markers IL‐18 and CTGF were increased as compared to normal weight individuals. However, no significant changes were observed in proteinuria or albuminuria.ConclusionsOur results indicate that augmentation of uAGT precedes albuminuria in young human adults with overweight and mice during the development of HFD‐induced T2D. These findings emphasize the relevance of uAGT and indicator of intrarenal/intratubular RAS activations and its potential as early biomarker of kidney injury during metabolic diseases as obesity and diabetes.
CUIDAME: a pharmacology and nutrition education program for ESPOL workers with Chronic Illnesses
Authors: Orellana‐Manzano* Andrea , Munoz‐Yee Victoria , Vega‐Falconez Noelia , Vizcaino Maria J. 
Published: May 2022

BackgroundPolypharmacy and self‐medication are frequent problems in patients with chronic non‐communicable diseases (CNCD). Accompanied by low health literacy (HA) affects the patient's medical treatment and nutritional status. For this reason, the present study aimed to design a virtual program of good practices for the consumption of medicines and food through digital media to improve the knowledge of these practices in workers of the Escuela Superior Politécnica del Litoral (ESPOL) with NCDs.MethodThe study was descriptive, observational, cross‐sectional, where the Ultra Indispensable Food Care Program for Efficient Medication (CUIDAME) was implemented. Twenty subjects from the areas of administration, teaching, and laboratory technicians from ESPOL participated. These had one or more NCDs and consumed more medications, supplements, or natural products. Updated and personalized information was provided to the educational needs through a Knowledge, Attitudes, and Practices (KAP) questionnaire. The educational material consisted of 4 sets of infographics, two online videos, and one digital guide. With a final survey, knowledge and level of satisfaction were assessed. For the statistical analysis, the InfoStat software was used.ResultsWe determined that the participants did not know about their treatment consumption and performed self‐medication. 87% of the participants had not received information on food for medical treatment, 48% had not yet made changes in their diet, but 65% considered that they could improve practices in consuming medications. After the program, it was shown that 53% had adequate knowledge about general dietary recommendations. In general, 59% reflected an excellent, and 41% reflected a very good total rating of the programConclusionInsufficient information about their nutritional therapy and forgetfulness in the medication intake schedule were the leading causes of poor knowledge and lack of treatment adherence. The best strategy for disseminating nutritional and pharmacological information was the CUIDAME Program and the guide given in virtual mode.
Effects of Creatine Nitrate and Caffeine Alone and Co‐ingested on Anaerobic and Muscular Endurance Performance
Authors: Koozehchian Majid S. , Mabrey Gina , Newton Andrew T. , Forbes Scott C. , Naderi Alireza , Haddad Monoem 
Published: May 2022

BACKGROUNDIngestion of creatine nitrate and caffeine individually improve anaerobic and muscular endurance performance; however, co‐ingestion of creatine nitrate and caffeine remains unclear.METHODSIn a double‐blind, randomized, placebo‐controlled cross over study, 12 resistance‐trained participants (22±1 y; 180.0±9.5 cm; 85.9±15.1 kg) completed 4 conditions in random order: creatine nitrate (CN: 5 g/day + 0.675 g/day of maltodextrin), caffeine (CAF: 400 mg/day + 5 g/day of maltodextrin), CN+CAF (5 g/day of creatine nitrate + 400 mg/day of caffeine), and placebo (PL: 5.4 g/day of maltodextrin). Supplements were ingested for 7 days with a 7‐day washout period between conditions. Before and after supplementation, muscular endurance (bench press and leg press; 3 sets at 70% 1RM) and a Wingate anaerobic test were performed. Peak power (PP), mean power (MP), and fatigue index (FI) were calculated. Data were analyzed using a two‐way (within‐within) analysis of variance (p≤0.05) with a Tukey’s post hoc when appropriate. Absolute change (post minus pre‐testing) was calculated and analyzed with a repeated‐measures ANOVA.RESULTSThere was a significant main effect of time for both leg press (p=0.009) and bench press (p<0.001) with post‐testing greater than pre‐testing; however, there were no differences between conditions. For peak power, there was no significant interaction (p=0.436), the main effect of time (p=0.967), or differences between conditions for absolute change (p=0.436). For mean power, there was a significant interaction (p=0.037); however, post hoc analysis revealed no significant pairwise effects; however, absolute change for mean power between conditions was significant (p=0.037), with post hoc analysis revealing a trend (p<0.10) for both CN conditions (CN alone and CN+CAF) to be superior compared to CAF. There were no significant differences between conditions or overtime for fatigue index.CONCLUSIONShort‐term CN and CAF ingested either individually, or co‐ingested could not augment upper or lower body muscular endurance or anaerobic performance compared to PL in resistance‐trained participants.