bluuebunny/crossref_metadata_2025_split huggingface dataset:
Catestatin regulates core bioenergetic and metabolic functions of the myocardium
Authors: Mahata Sushil K. , Mahata Sumana , Pasqua Teresa , Avolio Ennio , Tang Kechun , Bandyopadhyay Gautam , Webster Nicholas J. G. 
Published: May 2022

ObjectiveHypertension is a global pandemic, affecting more than one billion people. We have recently reported that chromogranin A (CgA) peptide catestatin (CST: human CgA352‐372) decreases blood pressure in hypertensive CST knockout (CST‐KO) mice by inhibiting catecholamine secretion and decreasing inflammation. Using CST‐KO mice we show here that CST regulates core bioenergetic and metabolic functions of the myocardium.Methods and ResultsWe carried out transcriptomic analyses (gene microarray) of the left ventricles of wild‐type (WT) and CST‐KO mice. Analysis revealed altered expression. Downregulated genes, which were most enriched in pathways that impact muscle conduction and contractility include, among other genes, Glo1 (glyoxylase 1). Upregulated genes include, among others, Gm7120 (predicted gene 7120), Astn2 (astrotactin 2), and Defa20 (defensin a‐20). We validated several of these up‐ and downregulated genes by qPCR and found that CST supplementation reversed these aberrant expressions in all cases. Targeted qPCR analysis of sarcomeric genes revealed reduced expression of Tnni3 (cardiac troponin I), Tnnt2 (cardiac troponin T), Myl4(atrial light chain‐1) and Myl7 (atrial light chain‐2) genes in CST‐KO mice. CST treatment increased expression of Tnni3, Tnnt2, Actc1 (cardiac a‐actin), Myl4 and Myl7 in both WT and CST‐KO mice albeit to a much higher level in CST‐KO mice. An analysis of mitochondrial genes revealed reduced expression of Ndufa3 (complex I), Sdhc (complex II) and Atp5j (complex V) genes in CST‐KO mice. CST treatment increased the expression of all three genes in WT and CST‐KO mice. Taken together, these unbiased approaches and qPCR validation studies provide insight into how an inflamed dysfunctional heart in the absence of CST may impact muscle function (contractility), rhythm, mitochondrial bioenergetics, and conduction.CST‐KO mice showed decreased uptake of 2‐deoxy glucose in the left ventricle compared to WT mice. Furthermore, glucose‐6‐phosphate utilization was significantly lower in CST‐KO compared to WT mice. We also found that compared to WT mice, CST‐KO mice show a significantly higher uptake of fatty acids. Furthermore, 14C‐labeled acid soluble metabolites were significantly higher in CST‐KO compared to WT mice, suggesting that increased dependence on fatty acids for energy needs make CST‐KO hearts resistant to insulin.ConclusionsTaken together with the cardiometabolic shifts (decreased glucose utilization with a concomitant increase in the utilization of fatty acids), these findings indicate that CST maintains cardiometabolic homeostasis; it does so at least in part by acting as an insulin‐sensitizing peptide which regulates myocardial choice of substrates for generating energy (i.e., ATP).
Chromogranin A plays a crucial role in age‐related development of insulin resistance and hypertension
Authors: Mahata Sushil K. , Liu Matthew A. , Jati Suborno , Tang Kechun , Bogaart Geert , Ghosh Gourisankar 
Published: May 2022

ObjectiveAging is a public health, societal, and economic problem around the world. Adults aged 65 years and older are the fastest growing segment of the US population, and their number is expected to double to 89 million by 2050. Two physiological processes that are more prevalent in older populations are hypertension and diabetes. Chromogranin A (protein: CgA; mouse gene: Chga) knockout (Chga‐KO) mice are associated with hypertension but also protected against developing autoimmune diabetes; however, the effect of these phenotypes on mice as they age including mortality remains unclear, as does a possible mechanism for developing both phenotypes. This study examined the long‐term effects of aging in Chga‐KO mice and sought to elucidate a possible mechanism for both hypertension and diabetes by focusing on the inflammatory pathway.Methods and ResultsChga‐KO mice display two opposite aging phenotypes: hypertension but heightened insulin sensitivity. We determined these phenotypes in aging CgA mice by measuring blood pressure and blood glucose. In comparison, aging wild‐type (WT) mice gradually lost glucose tolerance and insulin sensitivity. Moreover, while aging WT mice had increased inflammation with higher plasma TNF‐a, IFN‐g and CCL2 and increased mitochondrial fission, these phenotypes were the opposite in aging Chga‐KO mice. Chga‐KO mice also showed increased expression of mitochondrial and nuclear‐encoded complex I genes, implying that they were healthier than WT mice. Most intriguingly, the hypertension in Chga‐KO mice spontaneously reversed with aging. Supplementation of Chga‐KO mice with pancreastatin (human CgA273‐301), a hyperglycemic peptide produced from CgA by proteolysis, increased both blood glucose levels and blood pressure, implicating hyperglycemia, and hypertension.ConclusionsWe conclude that age‐related insulin resistance and hypertension are caused in part by CgA.
Trichinosis of Marine Mammals in the Territory of Chukotka
Authors: Mashkina Darja , Bukina Lidiya , Gaponova Viktoriya 
Published: May 2022

Chukotka is the only region of the Russian Federation where traditional fishing of marine mammals, including the gray whale Eschrichtius gibbosus, is conducted. More than 70% of the indigenous population of the coastal settlements of the Chukchi Peninsula eat the meat of marine mammals. Due to the huge distances between communities, lack of infrastructure, limited access to medical and veterinary care, isolated studies of marine mammal infestation with trichinella larvae are being conducted.For the first time, trichinella in the Pacific walrus Odobenus rosmarus divergens in Chukotka were recorded in the coastal zone of the Bering and Chukchi Seas in the 1970s. In the future, trichinosis has not been studied in this region, which confirms the relevance of our work. Infection of marine mammals was studied by trichinelloscopy (compressor and biochemical). Bioassays were collected during fishing by sea St. John's wort in the village of Lorino. Infection rates were determined. Studies conducted on the Chukchi Peninsula confirmed the infection with trichinosis of the Pacific walrus and revealed the larvae of trichinella in lakhtak Erignathus b.nauticus and ringed seal Phoca hispida, and later in larga Phoca largha. The extent of invasion varied from 0.9% in larga to 4.3% in lakhtak. The gray whale is the most popular food item in the diet of the indigenous inhabitants of Chukotka. According to the results of the research, out of 119 individuals of gray whales, trichinella were not found. Given the trophic‐chorological connections of marine mammals, it is difficult to imagine a way of transmission of trichinella in marine ecosystems. However, previously obtained research results allow us to do this. On the Arctic coasts, especially near settlements, there are a large number of corpses of dogs, cats, rats, voles, carcasses of small seals that fall into marine ecosystems. Invertebrates, their adult and larval forms, as well as vertebrates ‐ fish and birds ‐ occupy an important place in the process of disposal of corpses. Studies have shown that various types of invertebrates from 1 to 59 hours, and vertebrates from 28 to 95 hours can be direct or indirect sources of the pathogen of invasion and contribute to their mechanical transfer. Thus, in the conditions of coastal Arctic marine ecosystems, successful circulation of trichinella is carried out, thanks to human economic activity and the specifics of the trophic‐horological relationships of obligate and potential host species. Therefore, monitoring of trichinella invasion in these regions is vital.
Intracellular Sphingosine‐1‐Phosphate is Involved in Lung Cancer Cells Proliferation
Authors: Terlizzi Michela , Colarusso Chiara , Pinto Aldo , Sorrentino Rosalinda 
Published: May 2022

BackgroundSphingosine‐1‐phosphate (S1P) is a membrane‐derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1‐5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive.ObjectivesThe aim of our study was to understand the role of S1P in healthy versus lung adenocarcinoma cells after Toll‐Like Receptors (TLRs) activation, well‐known modulators of sphingolipid metabolism.MethodsWe stimulated healthy fibroblast and lung adenocarcinoma cells with TLRs ligands and S1P, to compare physiological vs pathological S1P role. Pro‐inflammatory cytokines release, proliferation rate and the expression/activity of the mediators involved in the S1P signaling were evaluated.ResultsWe found that the activation of TLR9, but not of TLR4, was involved in the generation of S1P by means of ceramidase and sphingosine kinase I/II (SPHK I/II) activation in tumor cells but not in non‐cancerous cells. Through an inside‐out signaling, newly formed S1P was extruded from the cells through its transporter SPNS2, supporting TLR9/NF‐kB‐mediated TNF‐α release mainly via membrane S1PR3. These results were not observed in healthy fibroblasts, implying that this pathway was mainly involved in pathological conditions. Moreover, we found that S1PR3 and SPHK II were overexpressed in the nucleus of lung adenocarcinoma cells and that were associated to S1P‐mediated proliferation of tumor cells.ConclusionThese results prove that the endogenous TLR9‐induced S1P can on one side favor pro‐inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis‐associated inflammatory processes.
PAR2, not PAR1, regulates endothelium‐dependent vascular tone in resistance arteries
Authors: Zhang Xun , Lee Matthew D. , Buckley Charlotte , Wilson Calum , Hollenberg Morley , McCarron John G. 
Published: May 2022

Protease activated receptors (PARs), a subfamily of G‐protein‐coupled receptors, are activated by cleavage of its extracellular domains. Four mammalian PAR members have been identified (PAR1‐4). PAR1 is the thrombin receptor, while PAR2 is activated physiologically by peptides or N‐terminal proteolytic cleavage. The resulting cleaved N‐termini acts as a tethered ligand that initiates receptor signaling. PARs may regulate vascular tone, cell permeability and cell proliferation. However, the precise roles of PAR1 and PAR2 in regulating vascular tone via the endothelium, and how PAR‐evoked signals transmit extracellularly and intracellularly, are incompletely understood.To explore the physiological functions, and the underlying signalling pathways in native endothelium, PAR1 (TFLLR‐NH2) and PAR2 (2‐Furoyl‐LIGRLO‐amide; 2fLI) activating peptides were used, and changes in intracellular Ca2+ and contractility measured in en face rat mesenteric arteries. Surprisingly, PAR1 activation (TFLLR‐NH2or trypsin) neither triggered Ca2+ signals in endothelial cells or alter vascular tone. On the other hand, PAR2 activation (2fLI) evoked large global propagating Ca2+ waves in endothelial cells and endothelium‐dependent dilation of mesenteric arteries. The Ca2+ signals evoked by PAR2 activation were kinetically distinctive when compared to those of muscarinic receptor activation. PAR2 activation (2fLI) also evoked responses in different clusters of endothelial cells from those of muscarinic receptor (acetylcholine) activation. PAR2 activation with 2fLI or trypsin failed to evoke Ca2+ signals after depletion of the internal store with the Ca2+‐ATPase (SERCA) inhibitor, CPA. This result suggests that PAR2 evokes Ca2+ release from internal stores. Pharmacological block of inositol 1,4,5‐trisphosphate (IP3) with either 2‐APB or caffeine, also blocked PAR2‐evoked Ca2+ signals. Phospholipase C inhibition (U73122) also blocked the Ca2+ signals evoked by PAR2 activation. Therefore, PAR2 is a PLC linked G protein coupled receptor which evokes Ca2+ release from the internal store via IP3 receptors in endothelial cells. The TRPC3 blocker, PYR3 and store‐operated Ca2+ entry (SOCE) blocker, YM58483 each significantly supressed PAR2 mediated Ca2+ signals, suggesting internal store was replenished via TRPC3 channel and SOCE.In conclusion, PAR2 rather than PAR1, regulates vascular tone in mesenteric arteries by evoking Ca2+ release from the internal store via IP3 receptors to generate propagating Ca2+ waves in distinct sub‐populations of endothelial cells.
Validation of a new assessment tool for chest tube insertion on Thiel embalmed human cadavers: a feasibility study
Authors: De Mol Leander , Willaert Wouter 
Published: May 2022

IntroThiel embalmed human cadavers are considered as high‐fidelity simulator for several surgical procedures, including Chest Tube Insertion (CTI). In proficiency‐based education, targeted feedback is essential in order to improve learners’ technical skills. Assessments must be validated prior to their use. Validity can be established using Messick’s framework, which collects evidence from five sources. The ACTION tool is a new assessment tool, and consists of a procedure‐specific Rating Scale and an error checklist. It has been developed by a Delphi panel, which ensures content evidence. This study evaluates the feasibility of a validation study for the ACTION tool on Thiel embalmed human cadavers.MethodsA convenience sample of three medical undergraduates and one PhD candidate was recruited. All provided informed consent and demographic information. Each participant was scored and recorded while performing a CTI. Each recording was later scored by three raters. Rater agreement was evaluated by calculating a two‐way mixed effects, absolute agreement, single unit of measurement intraclass correlation coefficient (ICC) model. Internal consistency of the assessment was evaluated using Cronbach Alfa. Feasibility was evaluated using structured questions regarding recruitment and data outcomes.ResultsRecruitment capability and data collection procedures were evaluated as being feasible and suitable for the target study participants. All necessary resources are available and data analysis can be carried out.Preliminary results for the procedural steps indicate an ICC of 0.62, indicating moderate rater agreement, and a Cronbach Alfa of 0.94, indicating excellent internal consistency. For the errors, an ICC of 0.63, and a Cronbach Alfa of 0.68 was observed.ConclusionValidation of the ACTION tool following Messick’s framework is feasible. A rater training session can increase reliability. After validation, the assessment tool can be used to evaluate learners’ proficiency in CTI on Thiel embalmed human cadavers, prior to patient contact.
Gentamicin induces calciuresis by blocking TRPV5
Authors: van Megen Wouter H. , Beggs Megan R. , An Sung‐Wan , Ferreira Patrícia G. , Lee Justin J. , Wolf Matthias T. , Alexander R. T. , Dimke Henrik 
Published: May 2022

BackgroundTreatment with the aminoglycoside antibiotic gentamicin can be associated with severe side effects including renal Ca2+wasting. The underlying mechanism is unknown but proposed to involve activation of the Ca2+‐sensing receptor (CaSR) in the thick ascending limb (TAL), which would increase expression of claudin‐14 (CLDN14) and limit Ca2+ reabsorption. However, no direct evidence for this hypothesis has been presented.MethodsThe effect of gentamicin on renal Ca2+ handling was studied in vivo using mouse models with impaired Ca2+ reabsorption in the proximal tubule and TAL. The effects of gentamicin in CaSR activation were studied in vitro using a Cldn14 promoter luciferase‐reporter assay, while the effect of gentamicin on the distal nephron Ca2+ channel transient potential receptor vanilloid 5 (TPRV5) activity was determined by patch‐clamp in HEK293 cells.ResultsGentamicin increased urinary Ca2+ excretion in wildtype mice following acute and chronic administration. This calciuretic effect was unaltered in mice with genetic CaSR overactivation and remained present in furosemide‐treated animals. Moreover, gentamicin‐induced calciuresis was not significantly different in Cldn14‐/‐ mice or mice with impaired proximal tubular Ca2+reabsorption (Cldn2‐/‐ mice), compared to wildtype animals. In vitro, gentamicin failed to activate the CaSR. In contrast, patch‐clamp analysis revealed that gentamicin inhibited rabbit and human TRPV5 channel currents in a dose‐dependent manner. In accordance with a direct effect of gentamicin on TRPV5, chronic gentamicin administration downregulated gene expression of distal nephron Ca2+ transporters in mice.ConclusionWe show here that gentamicin does not cause hypercalciuria via activation of the CaSR‐CLDN14 pathway or by interfering with proximal tubular CLDN2‐dependent Ca2+ reabsorption. Instead, gentamicin blocks distal Ca2+ reabsorption by direct inhibition of the Ca2+ channel TRPV5. These findings offer new insights into Ca2+ wasting in patients treated with gentamicin.
Electronic Nicotine Delivery Systems Exhibit LowerToxicity Compared to Cigarettes: “The Replica Study Experience”
Authors: Distefano Alfio , Caruso Massimo , Emma Rosalia , Rust Sonja , Poulas Konstantinos , Zadjali Fahad , Boffo Silvia , Volarevic Vladislav , Mesiakaris Konstantinos , Karanasios Georgios , Al Tobi Mohammed , Albalushi Najwa , Giordano Antonio , Canciello Angelo , Arsenijevic Aleksandar , Ilic Aleksandar , Caruso Tancredi , Carota Giuseppe , Spampinato Maria R. , Zuccarello Pietro , Ferrante Margherita , Polosa Riccardo , Li Volti Giovanni 
Published: May 2022

Electronic nicotine delivery systems (ENDS) can reduce the health risks associated with chronic smoke exposure, and their potential benefits are the subject of intense scientific debate. The international “Replica Study Group” replicated independently three relevant studies from the tobacco industry on the cytotoxic and inflammatory effects of cigarette smoke and ENDS aerosol. Our primary goal was to establish the reliability of the results and the robustness of the conclusions.In order to assess cytotoxicity of smoke and aerosol, we exposed human bronchial epithelial cell (H292) to cigarette smoke and to ENDS aerosol at air‐liquid interface (ALI). Moreover, we aimed to assess different inflammatory and remodeling mediators release (IL‐6, IL‐8 and MMP‐1) from cells exposed to whole smoke (WS) and to smoke deprived of total particulate matter (vapor phase; VP).We were able to replicate the results obtained in the original studies on cytotoxicity confirming that almost 80% of the cytotoxic effect of smoke is due to the vapor phase of smoke. Moreover, our results substantiated the significantly reduced cytotoxic effects of ENDS aerosol vs. cigarette smoke. However, our data were notably different in terms of inflammatory and remodeling activity triggered by smoke.Altogether, the data obtained independently in different laboratories clearly confirm the reduced toxicity of ENDS products compared to smoke, thus providing a valuable tool to the harm reduction strategies in smokers. Otherwise, we were not able to replicate the results on inflammatory and remodeling mediators released from smoke exposed cells. This could be due to the lack of normalization of the quantity of cytokines released, with the number of viable cells, in the original paper, since the production of cytokines itself requires the metabolic capability of cells.
Hypoxia Decreases Jump Rate and Increases Lactate Production in Late‐Stage Grasshoppers
Authors: Heiting Grace M. , Vanasse Sarah B. , Kirkton Scott D. 
Published: May 2022

Insects are the most evolutionarily successful animals in part due to their air‐filled tracheal respiratory system. The tracheal system is an intricate network of tubes that exchange gases between the atmosphere and tissues. Some insects, including grasshoppers, also have air sacs that enhance convective airflow. During intermolt periods, the American locust (Schistocerca americana) doubles their body mass, which may compress the tracheae. In a recent study, late‐stage grasshoppers had reduced air sac ventilation frequency, decreased convective capabilities, and less of their body dedicated to their respiratory system. We hypothesized that these developmental changes may reduce both oxygen delivery and jump performance. We forced early‐stage (day 2) and late‐stage (day 8) 6th instar grasshoppers to jump in artificial oxygen atmospheres (5%, 10%, 21%, 40% oxygen: balance nitrogen) in a 100 L gloved box. After jumping, grasshoppers were immediately frozen in liquid nitrogen for a fluorometric lactate assay. We predicted that if oxygen delivery is compromised during development, then late‐stage grasshopper jump rate and lactate production would correlate with oxygen level.Within the first one to two minutes of jumping, there were no significant differences in jump rate between early‐ and late‐stage grasshoppers at any oxygen level. This result was not surprising since previous work has shown that the first two minutes of jumping is fueled primarily by anaerobic ATP production. At moderate hypoxia (10% O2) and normoxia (21% O2), there were no differences in jump rate between early‐ and late‐stage grasshoppers. In extreme hypoxia (5% O2), there was no difference in jump performance for the first two minutes, but late‐stage grasshoppers had a 1.6‐fold lower average jump rate in minutes three through five. In hyperoxia (40% O2), jump performance was similar during the first minute, and late‐instar grasshoppers jump rates increased on average by 1.4‐fold in minutes two through five.Late‐instar grasshopper muscles produced significantly greater lactate regardless of oxygen level. Within each group, oxygen level did not significantly affect jumping muscle lactate production. However, late‐stage grasshoppers produced 26.3% significantly more lactate in 5% oxygen than 40% oxygen (post‐hoc Student’s t; p<0.04). Future research will examine how age and oxygen level impact HIF‐1 expression in grasshoppers. Since late‐stage grasshoppers have lower jump rates and increased lactate production, we predict they will also have elevated HIF‐1 protein levels.