bluuebunny/crossref_metadata_2025_split huggingface dataset:
Supraphysiological Levels of Testosterone Induce Cardiac Dysfunction via NLRP3 Inflammasome Activation in Macrophages
Authors: Alves Juliano V. , Costa Rafael M. , Omoto Ana C. M. , Oliveira‐Neto Jose T. , Silva Josiane F. , Machado Mirele R. , Rosa Ualter G. , Bonato Vania L. D. , Tostes Rita C. A. 
Published: May 2022

IntroductionTestosterone (Testo) modulates vascular tone and cardiac performance. Athletes who use Testo at supraphysiological doses exhibit increased blood pressure, higher inflammatory marker levels, vascular dysfunction, and cardiac hypertrophy. NLRP3 inflammasome activation, as part of the innate immune system response, contributes to proinflammatory cytokines production, leading e.g to cardiac dysfunction.HypothesisWe hypothesized that supraphysiological levels of Testo promotes cardiac dysfunction via generation of mitochondrial reactive oxygen species (mROS) and activation of the NLRP3 inflammasome.MethodsMale, 12 week‐old wild type (WT) and NLRP3 knockout (NLRP3‐/‐) mice were used. Mice were treated with testosterone propionate [Testo‐P (10 mg/kg)] or vehicle for 30 days. Cardiac function was evaluated using echocardiography. After in vivo experiments, western blot and ELISA were performed to evaluate NLRP3 inflammasome components. ROS generation was evaluated by lucigenin. Bone marrow‐derived macrophages (BMDMs) were isolated, primed with lipopolysaccharide (LPS [500 ng/mL 4 h]) and stimulated with Testo [10‐7 M], for 4, 6, 12 and 24 h. ROS generation was evaluated by DHE fluorescence and MitoSOX.ResultsEchocardiography showed cardiac dysfunction in WT mice treated with Testo‐P, characterized by reduced ejection fraction, shortening fraction, cardiac output and systolic volume. Testo‐P also increased interventricular septum, left ventricle posterior wall and decreased left ventricle internal diameter. All these effects were observed in NLRP3‐/‐mice. Furthermore, WT mice treated with Testo‐P showed increased cardiac expression of NLRP3 receptor, active Caspase‐1 and IL‐1β levels. These effects were prevented in NLRP3‐/‐. In addition, increased ROS generation was observed in the left ventricle of WT mice, but not NLRP3‐/‐ mice, treated with TP. Testo‐P‐treated WT mice showed increased macrophage infiltrate in the left ventricle, and this effect was not seen in the WT vehicle mice. In in vitro experiments, BMDMs primed with LPS and stimulated with Testo showed increased expression of the NLRP3 receptor and IL‐1β levels after 12 and 24 h. In addition, Testo‐stimulated BMDMs exhibited increased mROS generation.ConclusionSupraphysiological levels of testosterone induce cardiac dysfunction via mROS generation and NLRP3 inflammasome activation. This study was approved by the Ethics Committee on Animal Experimentation of the Ribeirao Preto Medical School (020/2021).
Dose dependent effects of green tea extracts in the skeletal development of a Down syndrome mouse model
Authors: Martínez Abadías Neus , Llambrich Sergi , González Rubèn , Albaigès Julia M. , Wouters Jens , Roldán Jorge , Salassa Sara , Van Bulck Vicky , Wouters Kaat , Callaerts‐Vegh Zsuzsanna , Sharpe James , Dierssen Mara , Vande Velde Greetje 
Published: May 2022

Down syndrome (DS) is a congenital disorder caused by trisomy of chromosome 21. The genetic imbalance results in intellectual disability, but also alters the development of different body systems, including the cranial and postcranial skeleton. To normalize altered development and to rescue skeletal phenotypes, green tea extracts enriched in EGCG (GTE‐EGCG) have been proposed for therapy. In this study, we longitudinally followed the skeletal postnatal development in a mouse model for DS, Ts65Dn, and assessed the effects of both a high dose and a low dose of pre‐ and postnatal GTE‐EGCG treatment. We administered a green tea extract containing 45% EGCG at doses of 100 or 30 mg/kg/day to pregnant Ts65Dn females, starting at embryonic day 9 and continued until adulthood. We longitudinally scanned wild‐type and Ts65Dn mice at postnatal days 3, 14 and 29 using in vivo micro‐computed tomography (µCT).Quantitative morphometric analyses showed that TS mice have a significantly different skull and mandibular shape than WT mice, as well as reduced bone mineral density (BMD) of the premaxilla and humerus bones that were detected already after birth. Moreover, Ts65Dn mice presented shorter faces, humerus and scapula, with significant differences appearing later in development, at the juvenile stage. Treatment with GTE‐EGCG generally deviated both WT and TS mice from normal growth trajectories, exacerbating some morphological anomalies, such as skull brachycephaly and reduced bone dimensions and mineral density. However, the effects of pre‐ and postnatal GTE‐EGCG administration were dose‐dependent, and low dose GTE‐EGCG treatment presented rescuing effects in facial shape. Our results underscore the complex nature of the DS phenotype and the need of further research to characterize the diverse effects of GTE‐EGCG treatment on different systems and structures and to fine‐tune dose and timing to achieve optimal safety and therapeutic potential.
Neutrophil Extracellular Traps Contribute to Perivascular Adipose Tissue Dysfunction in High‐Fat Diet Obese Mice
Authors: Costa Rafael M. , Neves Karla , Lopes Rhéure , Alves Juliano , Fedoce Aline , Silva Josiane , Rodrigues Daniel , Prado Douglas , Schnaider Ayda , Cunha Fernando , Alves José Carlos , Tostes Rita 
Published: May 2022

BackgroundObesity triggers functional changes in the perivascular adipose tissue (PVAT), favoring the release of vasoconstrictor factors and the activation of contractile mechanisms in vascular smooth muscle cells. In the PVAT of obese individuals, there is infiltration of immune cells, including neutrophils. A neutrophil defense mechanism is the formation of neutrophil extracellular traps (NETs), a cytotoxic framework that allows maintenance and expansion of inflammatory processes. We hypothesize that PVAT dysfunction in obese mice is due to increased neutrophil infiltration, NETs formation and inflammation.MethodsSix weeks‐old male Balb/C mice received control (CD) or high fat diet (HFD) for 18 weeks. Vascular function was assessed in endothelium‐intact mesenteric arteries in the presence or absence of PVAT. Mice were treated with DNAse type 1 (10 mg/kg for 14 days, every other day). Molecular mechanisms were investigated. Experiments were approved by the Ethics Committee on Animal Research of the Ribeirao Preto Medical School, University of Sao Paulo (protocol nº 209/2016)ResultsValues are expressed relatively to the contraction triggered by 120 mM KCl [Emax (% KCl)]. In Balb/C mice, the PVAT decreased the contractile response in control arteries [Control PVAT (‐): 123.2±8.1 n=6; Control PVAT (+): 64.4±4.0 n=6]. However, HFD promoted partial loss of PVAT anti‐contractile effect [HFD PVAT (‐): 127.9±6.9 n=6; HFD PVAT (+): 98.3±7.4 n=6]. In PVAT of obese mice there was a greater amount of neutrophils, compared to PVAT from control mice [Control: 8.0% n=5; HFD: 18.9% n=5]. In PVAT of obese mice there was greater formation of NETs, and production of interleukin 6 (IL‐6), compared to PVAT from control mice [NETs in pg/ml – Control: 30.5±2.7 n=6; HFD: 71.4±3.2 n=6; IL‐6 in pg/100 mg of PVAT – Control: 45.6±1.9 n=6; HFD: 98.4±3.1 n=6]. DNAse treatment reduced the formation of NETs and IL‐6 in the PVAT of obese mice [NETs in pg/ml – HFD+DNAse: 18.3±1.2 n=6; IL‐6 in pg/100 mg of PVAT – HFD: 48.1±3.4 n=6]. DNAse treatment restored PVAT anticontractile function in obese mice [HFD+DNAse PVAT (+): 71.3±7.4 n=6]. Adipose tissue of obese humans exhibited greater formation of NETs and production of IL‐6 compared to adipose tissue of eutrophic humans [NETs in pg/ml – Eutrophic: 20.5±3.7 n=7; Obese: 79.2±1.3 n=7; IL‐6 in pg/100 mg of adipose tissue – Eutrophic: 9.7±2.9 n=7; Obese: 27.7±2.6 n=7].ConclusionThese results indicate that, in obesity, there is a greater neutrophils infiltration in the PVAT and, consequently, greater formation of NETs and inflammation, events that contribute to PVAT dysfunction.
FTIR Spectroscopy Reveals Molecular Changes in BPC 157 Treated Blood Vessels
Authors: Gamulin Ozren , Krajacic Maria , Oroz Katarina , Coric Luka , Dretar Vilim , Strbe Sanja , Seiwerth Sven , Sikiric Predrag 
Published: May 2022

From the rats treated with stable gastric pentadecapeptide BPC 157 (10ng/kg ip) or saline (5ml/kg ip) at 5 min before sacrifice, the spectra of thoracic aorta tissue samples were compared. From each rat, 20 samples were cut and FTIR spectra were recorded.Vibrational spectra of the samples were recorded with a Perkin‐Elmer Spectrum GX spectrometer equipped with liquid N2 refrigerated Mercury Cadmium Telluride (MCT) detector. Data were acquired in 450 ‐ 4000 cm‐1 spectral range, in transmission mode with a resolution of 4 cm‐1.First, all recorded spectra were baseline corrected and normalized. Then we utilized the principal component analysis (PCA) to examine the possibility of separating BPC treated and control samples on a PCA score‐score graph. Using 100 spectra of BPC treated and 100 spectra of control samples, a PCA model was made. As seen in Figure 1, there is a clear distinction, mostly due to the PC1 score, between spectra of BPC treated (red) from the control group (green).To identify the parts of FTIR spectra that contributed the most to the separation between BPC treated and the control group of spectra differential spectrum was calculated and compared with PC1 loadings.Differential spectrum and PC loadings show that main bands responsible for separation of the BPC treated and control sample spectra are in the range from 930‐1200 cm‐1, then vibrational bands at 1660 cm‐1,1730 cm‐1, 1120 cm‐1 and in the range from 2840 – 3090 cm‐1. In the first range, there are three vibrational bands at 968 cm‐1, 1087 cm‐1 and 1240 cm‐1 that belong to P‐O symmetric and asymmetric vibrations in nucleic acids respectably, and vibrational band 1170 cm‐1 that belongs to C‐O and C‐O‐C stretching vibrations of carbohydrate. The vibrational band at 1660cm‐1 belongs to amid I stretching vibrations of proteins, and the last range of spectral lines belongs to C‐H symmetric and asymmetric C‐H, C‐H2 and C‐H3 vibrations in lipids. The line at 1730 cm‐1 that is very pronounced in control group spectra is C‐O symmetric vibration in fatty acids. Most prominent difference between BPC treated and control group was observed region of C‐H vibrations of lipids and vibrational line at 1730 cm‐1 of fatty acids.Thus, different FTIR spectra evidenced that BPC treatment causes changes of the tissue at molecular level.
Anatomical Study of Abdominal Wall Muscles Innervation
Authors: Alblowi Abdulrahman , Malikov Serguei , Perez Manuel , Labrousse Marc , Settembre Nicla 
Published: May 2022

BackgroundA variety of surgical specialties use different abdominal wall incisions to access the abdominal cavity. A thorough understanding of the anatomy, distribution, communication and course of these nerves are essential for surgeons. Anatomical knowledge remains limited. The anatomic relationships to other landmarks such as bone structures, ligaments, and muscles are important in prevention of peri‐operative complications such as abdominal muscle weakness, flank bulge, and incisional hernias. Imaging modalities do not provide many details about abdominal wall nerves.PurposeThe goal of this study is to provide a better understanding of the directions, course, communications, and branches of abdominal muscle wall nerves in order to minimize the risk of nerve injury.MethodFour frozen‐preserved cadavers with no abdominal wall scars visible were dissected layer by layer. The ninth, tenth, eleventh, and twelfth intercostal nerves were dissected from the midline to the posterior axillary line.ResultThe ninth, tenth, eleventh, and twelfth thoracolumbar nerves run in the same plane, between the internal oblique muscle and the transverse muscle, with varying directions and course. The T9 intercostal nerve has a horizontal course between the muscles, whereas the T10 and T11 and T12 have an oblique course toward the midline. Eleventh intercostal nerve, and twelfth subcostal nerves are bigger compared to the other nerves, with several communications and branches.ConclusionThe anatomy of nerves in abdominal muscles appears to be more complex and variable.Eleventh intercostal nerve and twelfth subcostal nerves appear to have larger contribution to the abdominal wall muscle innervation. They have a complex and variable network of communication. Preserving these nerves during surgical incisions, operation, and closure could help in the prevention of postoperative abdominal wall complications.
Alpha‐ketoglutarate promotes goblet cell differentiation and alters metabolome in a mouse model of colitis
Authors: Bravo Iniguez Alejandro , Tian Qiyu , Du Min , Zhu Meijun 
Published: May 2022

BackgroundAlpha‐ketoglutarate (aKG) is a key intermediate of the tricarboxylic acid (TCA) cycle. Previously, we demonstrated that aKG supplementation mitigates inflammation, enhances epithelial barrier integrity, and promotes oxidative phosphorylation in dextran sulfate sodium (DSS)‐induced colitis of mice. Intestinal epithelial homeostasis is regulated by cell energetics and metabolites from the microbiome. Given the importance of the mucosal layer in maintaining gut epithelial barrier function and microbiome composition, we hypothesized that dietary aKG facilitates the differentiation of secretory lineage epithelial cells and improves the gut metabolome.MethodsC57BL/6 male mice received drinking water with or without 1% aKG for three weeks, when mice were subjected to DSS induction for 7 days followed by 7 days recovery. The GC‐MS analysis was used to profile the cecal metabolomic changes due to aKG supplementation. In addition, colonic tissues were sampled for both biological and histochemical analyses.ResultsGC‐MS metabolomics revealed separation of the cecal metabolites between control (CON) and aKG supplemented (aKG) mice. In total, the contents of 40 metabolites were found to be different between CON and aKG groups. These metabolites include amino acids, fatty acids, and the B vitamin, niacin, succinic acid (butanedioic acid), the conjugate acid of succinate. Putrescine, a polyamine produced by intestinal microbiota and methysuccinic acid were among the metabolites with the largest fold change increased by dietary aKG. Amino acids including essential amino acids and those involved in the TCA cycle such as threonine and aspartic acid were elevated in the aKG group, of which, glycine displayed a 10‐fold increase. On the other hand, ornithine, an intermediate of the urea cycle, and amide products such as oleamide and urea were significantly decreased in aKG group. Pathway analysis revealed that the urea cycle, along with and arginine and proline metabolism and glutathione metabolism were altered due to aKG supplementation. Furthermore, aKG supplementation increased goblet cells based on Alcian blue staining, associated with increased mRNA expression of Mucin 2 and goblet cell differentiation markers Trefoil factor 3 and Krüppel‐like factor 4.ConclusionTogether, dietary aKG enhances intestinal epithelial differentiation to favor goblet cell differentiation, which correlates with improvement in metabolome.
Endogenous NRG4 promotes ST3GAL4 expression, leukocyte infiltration, and inflammation in IL‐10R neutralization colitis
Authors: Bernard Jessica K. , Schumacher Michael , Liu Cambrian , Katada Kay , Frey Mark 
Published: May 2022

BackgroundThe ErbB family of receptor tyrosine kinases are essential for the regulation of colonic epithelial cell proliferation, differentiation, survival, and wound healing. We previously showed that ErbB4 is induced as a compensatory response by intestinal inflammation; treatment with exogenous NRG4 (an ErbB4 ligand) protects colonocytes against cytokine‐induced apoptosis and ameliorates murine experimental colitis. Exogenous NRG4 can further alleviate intestinal inflammation by suppressing the activity of pro‐inflammatory macrophages. However, the role of endogenous NRG4 in the intestine has not been described.HypothesisBased on our previous studies, we hypothesized that NRG4 deletion would exacerbate intestinal inflammation.MethodsColitis was induced in 5‐week‐old wild‐type (WT) and NRG4 knockout (KO) mice by i.p. injection with 1 mg/kg IL‐10R neutralizing antibody, weekly for four weeks. Inflammation was assessed by fecal lipocalin‐2 ELISA, plasma myeloperoxidase ELISA, and total spleen weight. Disease was assessed by histological scoring of paraffin embedded colon rolls. Cytokine (TNF, IL‐1b, IFNg, and IL‐6) expression was determined by qPCR on RNA from full thickness distal colon homogenates. ST3GAL4 expression was assessed by qPCR and immunohistochemistry. Intestinal immune cell populations were identified and analyzed by mass cytometry (CyTof) using FlowJo software.ResultsFecal lipocalin expression was elevated in all mice after IL‐10R antibody treatment. Surprisingly, WT mice displayed significantly more fecal lipocalin than NRG4 KO cage mates. Similarly, after 4 weeks, WT mice had significantly higher plasma myeloperoxidase (2‐fold, p<0.001), increased spleen weight (57% more, p<0.01), and more histological injury and inflammation (72% higher score, p<0.05) versus their KO cage mates. Furthermore, mRNA levels for IFNg (3.7‐fold difference, p<0.01) and IL‐6 (1.6‐fold difference, p<0.05) were significantly higher in WT versus KO cage mates. CyTof analysis identified a reduction in CD8+ T cell populations, as well as alterations in overall T cell populations. Expression of ST3GAL4, a sialyltransferase that contributes to inflammatory leukocyte recruitment, was significantly reduced in NRG4‐null colons and epithelia (>90% reduction, p<0.01 and p<0.001 respectively), suggesting a mechanism for altered T cell populations.ConclusionIn contrast to the protective effects of exogenous NRG4, endogenous NRG4 appears to participate in recruitment of CD8+ T cells and intestinal inflammation, possibly through regulation of ST3GAL4.
Impact of Smoking On Immunomodulation Of Dental Pulp Stem Cells
Authors: Tahrani Leyla , Vang David , Han Xiaoyuan , Xiao Nan 
Published: May 2022

COVID‐19 has caused a global pandemic since 2019. After infecting the host cells, SARS‐CoV‐2 shows ability to hijack the host immune system and launch a cytokines‐storm. Smoking is reported to predispose patients to the respiratory system damage and lead to increased vulnerability to SARS‐CoV‐2 and more severe outcomes. Stem cells (SCs) are proposed as an alternative therapy of COVID‐19, especially the severe cases.Objectiveto investigate the impact of smoking and vaping on the inflammatory response of dental pulp stem cells (DPSCs).Hypothesiscigarette smoking and vaping inhibit the immunomodulatory, antimicrobial, and regenerative properties of the DPSCs and will reduce the success of stem cell therapy of COVID‐19.MethodsDPSCs were treated with various doses of cigarette smoke condensate (CSC) or nicotine for 24‐96 hours. Cell lysate and concentrated cell culture medium were collected at the end of the treatment. Expression of inflammatory cytokines were determined using human cytokine array, ELISA and quantitative PCR.Resultsthe level of TGF‐b2□ TGF‐b3, pulmonary and activation‐regulated chemokine (PARC or CCL‐18), tissue inhibitor of metalloproteinases (TIMP)‐1 and LIGHT (TNF superfamily member 14) significantly decreased after 48 hours exposed to 100mg/ml CSC. Interestingly, nicotine induced a different pattern of change of cytokine release. ELISA and quantitative PCR also showed that CSC treatment led to elevation of pro‐inflammatory cytokine IL‐6 level and reduction of anti‐inflammatory cytokine TGF‐b1 level in DPSC.Conclusionour current results indicated that smoking changes the cytokine expression profile in DPSCS, which has been used in clinical trials in treating COVID‐19. In addition to sharing many similarities with bone marrow mesenchymal SCs, DPSCs are also easy to access and show low immunogenicity reaction, making them valuable source to battle COVID‐19. We will further investigate the mechanism of how the SCs attenuate the cytokine storm induced by COVID‐19.
A Mouse Model Tracking Cardiac Exosomes Spatially and Temporally
Authors: Chang Jiang , Luo Weijia , Dai Yuan , Chen Zhishi 
Published: May 2022

Exosomes are 30‐150 nm secreted extracellular vesicles containing DNA, RNA, and proteins with lipid bilayer membranes. They exist in most organs and body fluids, including blood, tears, urine, amniotic fluid, breast milk, etc. The profiles of DNA, RNA, and proteins contained in exosomes are distinct from the exosome‐releasing cells. Recent studies revealed that different pathological conditions or stimuli could alter exosome sorting preferences, which resulted in the changes of phenotypes or functions in exosome recipient cells or organs. They are emerging as a new form of messengers facilitating cross‐talk between cells, tissues, and organs. Therefore, a critical demand arises for developing a sensitive and non‐invasive tracking system for endogenous exosomes. Tracking the biodistribution of endogenous exosomes and kinetics of endogenous exosome release and uptake should significantly impact basic and translational studies of exosome‐mediated pathways.Based on the exosome membrane structure, two major strategies have been applied for exosome labeling: by a lipophilic reagent‐conjugated reporter that incorporates into the exosome membrane and by exosome surface marker fusion reporter proteins. However, several challenges to monitoring endogenous exosomes currently remain due to the limitation of these exosome‐labeling techniques.In this study, we chose Nano‐luciferase (NanoLuc) as a reporter. NanoLuc is more stable and smaller in molecular size (19 kD) and generates a 150‐fold stronger signal compared to traditional Firefly and Renilla luciferases. The half‐life of the NanoLuc luminescence signal is more than 2 hours, the longest amongst all known luciferases. Its ultra‐stable and highly sensitive signal makes it an ideal reporter for endogenous exosome labeling to achieve a safe, non‐invasive, and quantifiable exosome tracking objective.We exploited these NanoLuc properties by fusing it to the exosome surface marker CD63 for exosome labeling and generated a proof‐of‐concept mouse model to enable tracking of endogenous exosomes. To achieve spatial control of exosome labeling, CD63NanoLuc reporter expression was controlled by the cardiomyocyte‐specific αMHC promoter in the animal heart. Endogenous exosomes released from the transgenic mouse cardiomyocytes were labeled and tracked. Moreover, the loxP‐STOP‐loxP CD63NanoLuc cassette was introduced to achieve inducible expression of the CD63NanoLuc reporter in vivo upon tamoxifen treatment, allowing temporal control of exosome labeling and tracking and making such mice more versatile to satisfy diversified research goals. The specific labeling and tissue distribution of endogenous exosomes released from cardiomyocytes were demonstrated by luciferase assay and non‐invasive bioluminescent live imaging. Finally, when bred with gene‐specific floxed mice, this model can be used to study gene‐specific exosome functions. This endogenous exosome tracking mouse provides a valuable tool for a range of research applications.