bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impact of human pluripotent stem cell‐derived extracellular matrix proteins on cardiac cell fate decision
Authors: Holeiter Monika , Bluguermann Carolina , Evseenko Denis , Crooks Gay , Schenke‐Layland Katja 
Published: June 2021

Incomplete understanding of the various factors that drive cardiac cell fate decisions limits the development of clinically successful pluripotent cell‐based strategies for the diseased heart. The objective of this study was to identify crucial ECM proteins that drive cardiac differentiation, presuming that a healthy developing cardiac ECM may help regenerate damaged cardiac tissue by providing a suitable microenvironment for cell replacement strategies. We employed a modified cardiac differentiation protocol, which allowed the robust and reproducible generation of spontaneously beating embryoid bodies (EBs) (85% ± 17.6%) from human embryonic stem cells (hESCs). EB‐derived cardiomyocytes were characterized using immunohistochemistry (IHC) and flow cytometry (cardiac troponin T (cTnT) or sarcomeric myosin), as well as quantitative real‐time PCR (qPCR). Monitoring of cardiac‐specific ECM by IHC and qPCR, allowed the identification of an increased expression pattern of decorin and nidogen‐1 during the differentiation of pluripotent stem cells towards the cardiovascular lineage. We hypothesize that identifying cardiac‐specific ECM proteins secreted during cardiogenesis will lead towards the development of biomaterials that may allow regeneration of the ischemic heart.Grant Funding Source: Fraunhofer Gesellschaft
Dynamic Formation of the Chick Sympathetic Ganglia
Authors: Kulesa Paul , Kasemeier‐Kulesa Jennifer C. , Lefcort Frances 
Published: June 2021

The sympathetic nervous system (SNS) plays a vital role in maintaining homeostasis and is derived from the embryonic neural crest. Trunk neural crest cells migrate as individuals along a ventral pathway and aggregate into primary sympathetic ganglia (SG) that form as discrete structures along the vertebrate axis. Within hours of formation, sympathetic precursors migrate dorsally and form the permanent, secondary chain of SG.The cellular and molecular mechanisms that mediate dorsal migration and formation of the secondary SG are largely unknown. This is due to the ventral, less accessible location of this morphogenetic event deep within the embryo. Using transverse slice culture and confocal time‐lapse microscopy, we detail the events involved in chick secondary SG formation. We find that the primary SG move as a cohesive cluster during their dorsal migration. SG cells display extensive cell contacts among neighbors and project long filopodial extensions in the direction of travel towards the forming spinal nerve. Extensions from the spinal nerve interact with dorsally migrating SG cells in an exciting interplay of cell contact to position the secondary SG. I will discuss these cell behaviors and emerging molecular studies aimed at identifying the factors that drive proper SG positioning and development.
Sensory neurons accelerate reepithelialization through Substance P release in an innervated tissue‐engineered model of skin wound healing.
Authors: Blais Mathieu , Mottier Lorène , Cadau Sébastien , Bareil Rémi Parenteau‐ , Berthod François 
Published: June 2021

The keratinocytes are responsible for the restoration of the epidermal barrier through reepithelialization during the wound healing process in humans. The influence of sensory neurons on this mechanism is poorly understood. We tested whether sensory neurons influence reepithelialization via a secretion of the neuropeptide substance P (SP). We developed a new tissueengineered model of skin wound healing. It consists of a perforated tissue‐engineered skin made of keratinocytes expressing GFP and that was stacked on a dermis to reconstitute the wound bed. The dermal construct is endothelialized by human endothelial cells and innervated or not by mouse sensory neurons. The migration of keratinocytes was imaged and quantified with GFP fluorescence. Sensory neurons produce SP in the construct as shown with ELISA and immunofluorescence analysis. Keratinocytes were found to express the NK1 cell receptor for SP in the reconstructed epidermis. The rate of reephithelialization was increased in presence of sensory neurons and SP was shown to contribute to this effect using antagonist to NK1 and recombinant SP instead of neurons. We conclude that this new model is promising for the study of the influence of the peripheral nervous system on reepithelialization.Grant Funding Source: N/A
Intakes of total and plant protein are associated with greater muscle strength: The Framingham Osteoporosis Study
Authors: McLean Robert R , Hannan Marian T , Kiel Douglas P , Casey Virginia , Mangano Kelsey M , Sahni Shivani 
Published: June 2021

ObjectiveTo determine the relation of dietary protein with quadriceps muscle strength in Framingham Offspring participants.MethodsMultivariable regression assessed protein (g/d, from FFQ 1995–98) with isometric quadriceps strength measured by hand‐held dynamometer (kg, max of 2 trials) in 2,674 men & women. Least squares‐adjusted mean strength was calculated for quartiles of energy‐adjusted protein (residual method) and linear trend tested. Model 1 assessed total protein (TP) and model 2 assessed animal (AP) and plant (PP) proteins.ResultsMean age 60 ± 9y. Protein intakes (g/d) were TP: 78±27, AP: 54±22; PP: 24±9. Participants in high quartiles of TP (Q3–Q4) and PP (Q4) had higher quadriceps strength compared to those in Q1 (Table). No significant associations were seen for AP intake. Similar results were observed with protein as a continuous variable.ConclusionsTP & PP intakes, but not AP were positively associated with quadriceps strength; consistent with data showing that alkaline diets may favor the preservation of muscle mass in older adults.Funding source: NIH AR # 053205; AR/AG41398, Framingham Heart Study (N01‐HC‐25195) & General Mills Bell Institute of Health and Nutrition
Endothelial GATA6 deficiency promotes pulmonary arterial hypertension (PAH) through ER stress‐mediated CHOP upregulation
Authors: Chrobak Izabela Maria , Ghatnekar Angela , Lenna Stefania , van Deuren Rosanne , Radstake Timothy , Feghali‐Bostwick Carol , Trojanowska Maria 
Published: June 2021

PAH is characterized by pulmonary vasculopathy, pulmonary artery pressure and RV failure. Previous studies have implicated endoplasmic reticulum (ER) stress in several disorders, including atherosclerosis, but its involvement in PAH has not yet been investigated. GATA6 is expressed in quiescent vasculature, but is lost in vascular injury. Our goal was to investigate the role of endothelial GATA6 in mediating ER stress in PAH.Gata6 flox/flox mice were bred with mice expressing Cre under the ECs‐specific VE‐cad promoter (Gata6 CKO).Gata6 CKO mice developed PAH and had severe changes in hemodynamics under hypoxia. Furthermore, loss of GATA6 resulted in worsened vascular remodeling and RV hypertrophy in hypoxia. The ER stress‐related markers BiP, PERK, ATF4 and XBP were moderately upregulated, whereas pro‐apoptotic marker CHOP was significantly upregulated in Gata6CKO mice. Furthermore, upregulation of BiP protein was observed in Gata6CKO lungs by immunohistochemistry. Consistent with in vivo observations, depletion of GATA6 in HPAECs via siRNA resulted in upregulation of ER stress mediators including BiP, ATF4 and CHOP.Our findings suggest that GATA6 deficiency in endothelial cells leads to development of PAH, in part via upregulation of ER stress/CHOP pathway.This study was supported by NIH grant RO1 AR42334 and Postdoctoral Fellowships NIH training grant T32 AR 050958.
Surfactant activity and mobilization in minced saline extracts of neonatal brain, heart, kidney, liver, and lung
Authors: Shanklin Douglas Radford 
Published: June 2021

Minced saline tissue extracts yield minimum values higher than airspace washings, attributed to different proportions of saturated fatty acids and phosphatidylcholine, with typical hysteresis in both. Saline minced extracts from newborn autopsy tissues (Kimray Greenfield surfactometer) follow the formation of lung lesions; 16 newborns up to 90 hrs had a progressive but counter intuitive decline in lung minimum values, suggesting enhanced synthesis without therapy or recruitment of surfactant [Regression equation: minimum (dynes/cm) = −0.1936 (survival hrs) + 23.7040]. No difference was seen by gestational time (22–40 wks), as shown by Esterly (Am.J.Clin.Path. 46:649, 1966), a finding against surfactant deficiency due to prematurity per se. Lesser declines in the minimum was found in extracts of myocardium (43.1% of lung), kidney (17.8%), liver (16.5%), while the minimum worsened in extracts from cerebrum (+7.0%), the largest pool of phospholipids of the five tissues. This sequence suggests possible mobilization of surface active lipids during the first four days after birth, despite respiratory or other disorders. Twenty stillborns had similar results by gestational age; mean minimums for stillborn lung and liver were not significantly different from mean newborn values (lung, t = 0.392, p = 0.696; liver, t = 1.019, p = 0.315). Thus, newborn surfactant dynamics are systemic.Support: The Rhad Trust
Holliday Junction Resolvases: the Good, the Bad and the Ugly
Authors: West Stephen 
Published: June 2021

In simple organisms, Holliday junctions (HJs) that arise during recombination and the recombinational repair of DNA are resolved by a junction‐specific endonuclease activity. For example, in E. coli, the RuvC protein specifically recognises and binds HJs, and then promotes their cleavage by the introduction of symmetrically‐related nicks across the junction point. The resulting nicked duplexes can then be religated.In contrast, mitotic eukaryotic cells appear to have evolved a rather different mechanism of resolution, because in humans the removal of HJs is preferentially mediated by the BTR complex (BLM‐TopoIIIα‐ RMI1–RMI2), which combines DNA helicase and topoisomerase activities to bring about HJ ‘dissolution’. The benefit of this mechanism of HJ processing is that recombination events that link homologous chromosomes are guaranteed to be resolved to form non‐crossovers, thereby avoiding crossovers and the potential for loss of heterozygosity (LOH) of tumour suppressor genes ‐ a hallmark feature of cancer cells.Given the importance of avoiding crossovers, it is perhaps surprising that mitotic cells encode three junction‐specific nucleases that can resolve HJs to form crossovers. In humans, these are MUS81‐EME1, SLX1‐SLX4 and GEN1. How these nucleases are controlled, in order to avoid crossover formation, has revealed unexpected insights into mechanisms of Holliday junction processing.
A prospective study of global DNA methylation and development of adiposity in Colombian schoolchildren
Authors: Perng Wei , Mora‐Plazas Mercedes , Rozek Laura S. , Marin Constanza , Baylin Ana , Villamor Eduardo 
Published: June 2021

ObjectiveTo examine associations of global DNA methylation with changes in adiposity in a longitudinal study of school‐age children from Bogotá, Colombia.MethodsWe quantified methylation of LINE‐1 elements from peripheral leukocytes of 553 children aged 5–12 y at baseline using pyrosequencing technology (Qiagen). Anthropometric characteristics were measured periodically for a median of 30 months. We estimated mean change in 3 age‐standardized indicators of adiposity: body mass index (BMI) Z score (BMIZ), waist circumference Z score (WCZ), and triceps‐to‐subscapular skinfold thickness ratio Z score (SKFZ) according to quartiles of LINE‐1 methylation using multivariable mixed effects linear regression.ResultsThere were inverse, non‐linear relations of DNA methylation with BMIZ and SKFZ change in boys. After adjustment for baseline age and socioeconomic status, boys in the lowest quartile of LINE‐1 methylation experienced annual gains in BMIZ and SKFZ that were 0.06 Z/y (P=0.04) and 0.07 Z/y (P=0.03), respectively, higher than those in the upper three quartiles. There was an inverse, monotonic relation of LINE‐1 methylation with annual change in WCZ (P trend=0.02). DNA methylation was not related to development of adiposity during follow‐up among girls.ConclusionsGlobal DNA methylation was inversely related to development of central adiposity in boys.Grant Funding Source: This study was funded by the Robert Wood Johnson Foundation Research Project Awards. The Bogotá School Children Cohort is currently sponsored by the ASISA Research Fund at the University of Michigan.
γ‐Tocopherol additionally improves vascular endothelial function following smoking cessation by decreasing pro‐inflammatory responses
Authors: Mah Eunice , Ballard Kevin D , Pei Ruisong , Guo Yi , Volek Jeff S , Bruno Richard S 
Published: June 2021

Oxidative stress and inflammation that persist following smoking cessation limits the restoration of vascular endothelial function (VEF). We hypothesized that smoking cessation with γ‐tocopherol (γ‐T)‐rich supplementation would decrease oxidative stress and pro‐inflammatory responses and improve VEF beyond that of smoking cessation alone. A randomized, double‐blind, placebo‐controlled study was conducted in otherwise healthy smokers (≥10 cigarettes/d) who quit smoking for 7 d with placebo (n = 14) or γ‐T‐rich supplementation (n = 16; 500 mg γ‐T/d). Brachial artery flow‐mediated dilation (FMD), plasma cotinine, vitamin E, malondialdehyde (MDA), and pro‐inflammatory mediators were measured before and after 7 d of smoking cessation. Smoking cessation regardless of supplementation similarly decreased plasma cotinine whereas plasma γ‐T increased by 7‐fold (P<0.05) only in those receiving γ‐T‐rich supplements. Smoking cessation with γ‐T‐rich supplementation increased FMD responses beyond that of smoking cessation alone (P<0.05). Although MDA decreased similarly in both groups, myeloperoxidase and TNF‐α decreased only in those receiving γ‐T‐rich supplementation. These findings support that improvements in γ‐T status additionally improve VEF during smoking cessation likely by decreasing pro‐inflammatory responses. Funded by CT Department of Public Health.Grant Funding Source: Connecticut Department of Public Health
Diet‐induced obese mice exhibit heightened lung inflammatory and cross‐reactive CD8 T cell responses during a secondary 2009 pandemic H1N1 influenza infection
Authors: Milner J Justin , Sheridan Patricia A , Karlsson Erik A , Schultz‐Cherry Stacey , Shi Qing , Beck Melinda A 
Published: June 2021

Obese individuals were at greater risk for hospitalization and death following infection with the 2009 pandemic H1N1 influenza virus (pH1N1). Although cross‐reactive memory T cells can limit pH1N1 death and disease severity, we have previously shown obese mice and humans exhibit impaired memory T cell function. To determine if obesity results in greater pH1N1 morbidity as a consequence of impaired cross‐reactive T cell defenses, diet‐induced obese and lean, control mice were infected with a sublethal dose of influenza PR8. After 5 wk, the mice were challenged with a lethal dose of heterologous pH1N1. Despite a lack of differences in mortality, obese mice lost more weight and exhibited greater lung viral burden, immune cell infiltration, pathology and reduced lung integrity during the secondary challenge. Consistent with the elevated lung inflammatory responses, obese mice had a greater number of cytotoxic effector memory CD8+ T cells in the lungs. To investigate a potential mechanism for the excess inflammatory and pathological responses in the lungs of obese mice, we assessed the distribution of anti‐inflammatory regulatory T cells (Tregs). Although obese mice had twice has many lung Tregs compared with lean mice, Tregs isolated from obese were 40% less functional. Taken together, drastic impairments in Treg function may contribute to excessive inflammation and greater pH1N1 infection severity in the obese.Grant Funding Source: NORK DK56350
COCOA CONSUMPTION DOSE‐DEPENDENTLY IMPROVES FLOW‐MEDIATED DILATION AND ARTERIAL STIFFNESS IN HEALTHY SUBJECTS
Authors: Grassi Davide , Desideri Giovambattista , Necozione Stefano , Di Giosia Paolo , Cheli Paola , Barnabei Remo , Allegaert Leen , Bernaert Herwig , Ferri Claudio 
Published: June 2021

To investigate the effects of different doses of cocoa on endothelial function and arterial stiffness.According to a randomized, double‐blind, controlled, cross‐over design, 20 healthy volunteers were assigned to receive either five treatments with daily intake of 10 g cocoa (0, 80, 200, 500 and 800 mg cocoa flavonoids/day) in five periods lasting 1 week each.Cocoa dose‐dependently increased flow‐mediated dilation (FMD) from 6.2% (control) to 7.3, 7.6, 8.1 and 8.2% after the different flavonoid doses, respectively (p < 0.0001). Compared with control, even 80 mg cocoa flavonoids/day increased FMD (p < 0.0001). The change in FMD after 800 mg/day was not only significant compared with control (p < 0.0001) but also with 80 mg/day (p = 0.0003) and 200 mg/day (p = 0.05). Compared with control, carotid–femoral artery pulse wave velocity (PWV) dose‐dependently ( 0 mg vs. 200 mg: p = 0.0008; 0 mg vs. 500 mg: p < 0.0001; 0 mg vs. 800 mg: p < 0.0001) decreased.Our study showed for the first time that cocoa dose‐dependently improved FMD and decreased PWV. Our findings are clinically relevant, suggesting cocoa, with very low calorie intake, might be reasonably incorporated into a dietary approach representing a consistent tool in cardiovascular prevention. Starting from our findings, the European Food Safety Authority claimed a relationship between cocoa flavanols and endothelial‐dependent vasodilation.
Black soybean may be effective to recover the impaired insulin signaling in high cholesterol diet induced non‐alcoholic fatty liver disease mice model
Authors: Jung Ji‐Hye , Yoon Lee‐Na , Kim Hyun‐Sook 
Published: June 2021

NAFLD is strongly associated with insulin resistance and is defined by accumulation of liver fat more than 5% per liver weight. In this study, we investigated the effects of low and high concentrations of black soybean on the insulin signaling of mice fed high‐cholesterol diets. Mice were randomly allocated into 5 groups that were fed different diets: the normal cholesterol diets (NCD); high‐cholesterol diets (HCD); and HCD with 1%, and 4% black soybean powder (1B‐HCD, and 4B‐HCD, respectively). Blood glucose and insulin were significantly lower in the black soybean‐supplemented groups (129.2mg/dl, 1.76μIU/ml; 113.7mg/dl, 2.29μ IU/ml) than that in the HCD group (163.2mg/dl, 3.07μIU/ml). The serum HOMA‐IR and free fatty acid concentration were significantly higher in the HCD group than in the NCD, and it was decreased by black soybean supplementation. We measured adiponectin as it is critical adipokine involved in insulin resistance in NAFLD. The adiponectin was significantly lower in the HCD group than that in the NCD and significantly increased with black soybean supplementation. These results suggest that the black soybean restores the fasting glucose via adiponectin secretion.Grant Funding Source: ASN
Differential effect of mesenchymal stem cell therapy on cardiac glucose utilization in the infarcted heart following chow and high fat feeding
Authors: Hughey Curtis , Ma Lianli , James Freyja D , Bracy Deanna P , Wasserman David H , Rottman Jeffrey N , Shearer Jane 
Published: June 2021

OBJECTIVETo determine whether mesenchymal stem cell therapy minimizes abnormal cardiac glucose utilization following a myocardial infarction (MI) in the presence of dietary‐induced insulin resistance.METHODSC57BL6 mice were high fat‐fed (HFF) for eight weeks. Mice were separated into three groups: HFF‐SHAM, HFF‐MI+PBS in which mice were given a MI followed by intramyocardial injection of saline and the HFF‐MI+ MSC group receiving MSCs. HFF groups were age‐matched with chow‐fed mice.RESULTSFive weeks post‐MI, cardiac glucose utilization was evaluated in the conscious, unrestrained mouse by performing a hyperinsulinemic‐euglycemic (insulin) clamp coupled with [14C]‐2‐deoxyglucose administration. In chow‐fed mice, MSCs preserved glucose uptake (μg/mg/min) in the remote left ventricle (RLV) (7.36 vs. 5.16 vs 6.00; CHOWSHAM vs CHOW‐MI+PBS vs CHOW‐MI+MSC). In HFF mice, MSCs increased insulin‐stimulated glucose uptake in the periinfarct region (3.85 vs 3.84 vs 5.24; HFF‐SHAM vs HFF‐MI+ MSC vs HFF‐MI+MSC). RLV glucose uptake in the HFF‐MI+ MSC group was higher than that of the HFF‐MI+PBS mice (4.80 vs 3.38). Immunoblots indicate MSCs preserve insulin signaling (p‐Akt/Akt) in chow‐fed mice. In contrast, this ratio was similar in HFF groups.CONCLUSIONSMSC tberapy promotes insulin sensitivity post‐MI differently in response to a high fat diet. Funded by CIHR and Killam Trusts.
Human milk oligosaccharides reduce EPEC attachment in vitro and EPEC colonization in mice
Authors: Manthey Carolin Friederike , Autran Chloe , Eckmann Lars , Bode Lars 
Published: June 2021

Breastfeeding is associated with a lower risk for enteric infections, in part due to human milk oligosaccharides (HMO), unconjugated complex glycans that are present in human milk (5–15g/L), but not in infant formula. Enteropathogenic Escherichia coli (EPEC) are attaching/effacing pathogens that can cause enteric infections and serious diarrheal illness with potentially high mortality in infants, especially in developing countries. Our objective was to assess whether HMO reduce EPEC attachment in vitro and colonization in vivo.Physiological concentrations of HMO (10g/L) reduced EPEC attachment to cultured epithelial cells (HeLa and T84) by 99% compared to mock‐treated controls (p<0.0001). Galactooligosaccharides (GOS, 8g/L), currently added to infant formula, had no effect. In 7‐day old suckling mice, oral administration of HMO (15mg/d) thrice daily the day prior and after EPEC exposure reduced colon EPEC counts to 3.8E+06 compared to 1.1E+08 in PBS‐treated littermates (p=0.015) and 5.1E+08 in GOS‐treated littermates.Our data suggest that HMO reduce EPEC attachment in vitro and colon colonization in suckling mice. We now aim to identify which of the more than 130 different HMO contribute to the observed effects and whether our results translate to human infants.
Sustanon Administration Induces Satellite Cell Proliferation and Giant Fiber Formation in Growing Skeletal Muscle without Exercise
Authors: Allouh Mohammed Zohair , Aldirawi Mohammed Hasan 
Published: June 2021

Sustanon is one of the most commonly used anabolic androgenic drugs to increase skeletal muscle mass and strength. This drug is a blend of four esterized testosterone derivatives: testosterone propionate, testosterone phenylpropionate, testosterone isocaproate, and testosterone decanoate. Little is known about the effects of this drug on skeletal muscle at the cellular level. This study aimed to investigate the influence of Sustanon on the morphology of skeletal muscle fibers and the distribution of myogenic satellite cells (SCs) during postnatal growth. We hypothesized that Sustanon‐induced skeletal muscle hypertrophy is associated with an increase in the number of SCs. Robust immunocytochemical techniques and morphometric analyses were used to calculate the numbers of SCs and myonuclei within pectoralis muscle of the chicken. Sustanon significantly increased pectoralis mass and fiber size. All SC indices and the number of myonuclei increased significantly by Sustanon administration. This study indicates that Sustanon can induce hypertrophy and increases the numbers of SCs and myonuclei in skeletal muscle fibers without imposed exercise. Funds for this study were provided by a grant from the Deanship of Research at Jordan University of Science & Technology.Grant Funding Source: Jordan University of Science & Technology