bluuebunny/crossref_metadata_2025 huggingface dataset:
Cross‐tissue telomere length measurement to determine auxiliary marker of brain telomere length in neurosurgery patients
Authors: Lussier Stephanie , Braun Patricia , Potash James , Shinozaki Gen , Stevens Hanna
Published: November 2023

Background

Telomere length, the length of the chromosomal endcaps, is a commonly used biomarker of health and aging, as it may indicate the extent to which cells have undergone damage. Changes in telomere length have been connected to exposure to stress and poor physical health outcomes, such as heart disease, risk of stroke, and coronary artery disease. The use of telomere length as a biomarker, therefore, is reflective of its importance in cellular development and function. As a biomarker in human studies, telomere length is measured in cells taken from blood, saliva, or other easily obtained peripheral cells, even in the study of disorders of the central nervous system (CNS) such as psychiatric illness. It is unclear if telomere length plays a role in pathophysiological processes in the brain. This question could be better addressed by determining if telomere length changes in the CNS are commensurate with those in peripheral tissues.

Methods

In this study, we aim to look at the relationship between CNS neural telomere length measurements in comparison to peripheral measurements in two different tissue types to determine if peripheral telomere length measurements may be proxies for CNS measures. Samples of blood, saliva, and brain were taken in 22 neurosurgery patients before, during, and after surgery. Brain resection surgeries were performed to remove tissue at seizure focal points. Patients had an average age of 32.4 years ranging from 5 to 61 years old and 15 out of 22 patients (68%) were male. Telomere length was measured in all samples using quantitative PCR techniques.

Results

Blood telomere length, but not saliva or brain, negatively correlated with age (r=0.84, p<0.001), a previously reported phenomenon. Comparing pre‐ and post‐surgery samples, we found that saliva telomere length had a trend decrease (paired student's t‐test, p=0.08), while blood telomere length had no change. Intriguingly, we found a trend correlation between brain telomere length and pre‐surgery saliva telomere length (standardized beta = 0.47, r=0.47, p=0.09).

Conclusions

We found that telomere measures in saliva trended to be different before and after surgery, in which a significant intervening variable was use of synthetic glucocorticoids to prevent edema. We hypothesize that exposure to glucocorticoid even on this acute time course may affect saliva telomere length. Further, we have found that there may exist a relationship between brain and pre‐surgery salivary telomere length, but no other examined tissues (post‐surgery saliva, pre‐surgery blood, and post‐surgery blood). These findings may help to identify saliva telomere length as a potential correlate for brain telomere length, which may broaden its applications as a biomarker.

Support or Funding Information

University of Iowa CPH Advancing Graduate Student Success Award
Regulation of Cannabinoid‐mediated Neuroprotection by Hydrogen Sulfide in Bovine Isolated Retina
Authors: Mitchell Leah , Robinson Jenaye , Ngele Kalu , Opere Catherine , Ohia Sunny E , Njie‐Mbye Ya Fatou
Published: November 2023

Hydrogen sulfide (H2S) and endocannabinoids have been reported to possess neuroprotective properties in several mammalian tissues. In the present study, we will investigate the role of H2S in the neuroprotective actions of cannabinoids in the bovine isolated neural retina.

Methods

Isolated bovine retinae were exposed to oxidative stress using hydrogen peroxide (H2O2, 100 μM) for 10 minutes. Before exposure to stress, tissues were pretreated with inhibitors of H2S biosynthesis, amino‐oxyacetic acid (AOAA, 30 μM) and α‐keto‐butyric acid (KBA, 1 mM), and the CB1‐receptor antagonist, AM251 (100 nM) prior to treatment with methanandamide (1 nM – 100 nM). Lipid peroxidation was assessed using the Cayman 8‐isoprostane ELISA kit.

Results

In the presence of H2O2 (100 μM), there was a 20% increase in 8‐isoprostane levels when compared to control. At a concentration of 1 nM, methanandamide significantly (P<0.001) reversed the H2O2 (100 μM)‐induced augmentation in 8‐isoprostane levels in the neural retina. Interestingly, the neuroprotective action caused by methanandamide was abolished by KBA (1 mM) but not by AM251 (100 nM) or AOAA (30 μM).

Conclusions

Our results demonstrate that the neuroprotective action of methanadamide can be reversed by an inhibitor of the enzyme, 3‐mercaptopyruvate sulfurtransferase but not by the CB1‐receptor antagonist. Taken together, we conclude that the neuroprotective action of endocannabinoids involve the endogenous biosynthesis of H2S via the cysteine aminotransferase dependent pathway in the bovine retina.
Clinical evaluation of hop‐drug interactions using probe substrates of cytochrome P450 enzymes and a hop dietary supplement
Authors: Chen Luying , Tonsing‐Carter Alyssa , van Breemen Richard
Published: November 2023

Objective

The title states the study objective.

Method

Low doses of an oral cocktail of caffeine, tolbutamide, dextromethorphan, and alprazolam (probes for metabolism by CYP1A2, CYP2C9, CYP2D6, and CYP3A4, respectively) were administered to peri‐ and post‐menopausal women at baseline and then again after consuming a hop dietary supplement twice daily for 14 days. Serial blood samples were drawn and analyzed for concentrations of each probe substrate drug over time, and concentration‐time curve values were compared. A Shimadzu Nexera UHPLC system with a Shimadzu LCMS‐8060 triple quadrupole mass spectrometer was used with CID and SRM for the quantitative analysis of all four probe substrates in serum. Concentration‐time curves and other pharmacokinetics parameters were calculated using WinNonlin software.

Results

According to our preliminary in vitro data, hop preparations containing prenylated flavonoids and chalcones have the potential to inhibit some cytochrome P450 enzymes especially CYP1A2 and members of the CYP2C family, possibly contributing to prolonged drug half‐lives and elevated serum concentrations when co‐administered with specific therapeutic agents. To evaluate these safety issues in the clinic, we tested a botanically authenticated and chemically and biologically standardized hop dietary supplement in 16 peri‐ and post‐menopausal women (ages 40–75) for possible interactions with the pharmacokinetics of four FDA‐approved drugs (caffeine, tolbutamide, dextromethorphan, and alprazolam) as probe substrates for cytochrome P450 enzymes using ultra high‐pressure liquid chromatography/tandem mass spectrometry (UHPLC‐MS/MS). However, in this clinical evaluation, no changes in drug metabolism were detected after treatment with the hop dietary supplement.

Conclusion

Although in vitro assays had predicted in vivo drug‐botanical interactions, preliminary clinical data suggest no clinically relevant interactions of hop dietary supplements with drug metabolism.

Support or Funding Information

Funding was provided by the NIH Office of Dietary Supplements and the National Center for Complementary and Integrative Health (NCCIH) [P50 AT000155]
Using Gaming Strategies to Augment Pharmacology Content Retention in a Doctor of Pharmacy Program
Authors: Malhotra Ashim
Published: November 2023

Objective

1) To enhance learner engagement in acquisition of pharmaceutical science content at a three year pharmacy program using interactive games like Jeopardy! 2) Augment learner recall of fundamental concepts and information retrieval, targeting “cognitive functions” in Bloom's Taxonomy.

Methods

The Pacific University School of Pharmacy implements a three year curriculum where renal, immune, and cardiovascular pharmacology is taught in the P1 year. We implemented a gaming strategy to 1) capture learner interest, and enhance 2) participation and 3) recall by instructing sixteen groups of P1 students to prepare Jeopardy! question using poste lectures. Five selected groups were invited self‐select a “leader” and provided pressure‐sensitive buzzers. The remaining groups role‐played “Alex Trebek”, asking questions in a serialized format. Student perception was surveyed to determine whether the activity was 1) helpful, 2) repeatable, 3) important for memorizing content, and, 4) analyzing content for exam‐emphasis.

Results

We used Qualtrics to build and analyze data collected from our survey. Surprisingly, although 73% of the 57 respondents indicated that exercise was useful, 32% argued for conducting the activity outside of the class time. However, 77% suggested that the activity be repeated. Student showed improvement in performance when some of the game content was used for the final assessment.

Implications

This study suggested modification of gaming strategies for use in the pharmaceutical science curriculum, with either a shorter intervention, or one that is implemented in fewer blocks.
The Role of Nrf2 in Primary Human CD4 T Cell Activation and Differentiation
Authors: Turley Alexandra E , Zagorski Joseph , Rockwell Cheryl E
Published: November 2023

The transcription factor nuclear factor erythroid‐derived 2 like‐2 (Nrf2) responds to cell stress such as reactive oxygen species, electrophilic xenobiotics, and heavy metals. In response to such stress, Nrf2 induces a battery of detoxification, antioxidant, and metabolizing genes. In addition to its cytoprotective functions, Nrf2 has been implicated in immune modulation. Female Nrf2‐null mice develop a disease similar to systemic lupus erythematosus as they age. In addition, Nrf2‐null mice are more sensitive to a number of inflammatory stimuli such as cecal‐ligation and puncture and experimental autoimmune encephalomyelitis, and conversely, Nrf2 activation helps protect against these stimuli. Nrf2 has also been shown to modulate macrophage and dendritic cell function in vitro, and recently our lab has shown that Nrf2 skews CD4 T cell differentiation in primary mouse CD4 cells towards a Th2 phenotype. However, the function of Nrf2 in primary human CD4 T cell activation and differentiation remains unclear. Therefore, the purpose of these studies was to determine the role of Nrf2 in primary human CD4 T cell activation and differentiation. In these studies, peripheral blood mononuclear cells (PBMC) and CD4 cells were isolated from human blood. Cells were treated with either tert‐butylhydroquinone (tBHQ), a commonly used food additive and Nrf2 activator, or 1[2‐Cyano‐3,12‐dioxooleana‐1,9(11)‐dien‐28‐oyl]imidazole (CDDO‐IM), another Nrf2 activator, for 30min, then activated with anti‐CD3/anti‐CD28. Treatment of PBMCs with tBHQ for 6h increased expression of the Nrf2 target genes HMOX, GCLC, and NQO1. Both tBHQ and CDDO‐IM inhibited events of T cell activation at 24h, including production of the cytokines interleukin‐2 and interferon‐γ and expression of the cell surface markers CD25 and CD69. Isolated CD4 T cells show greater inhibition of early events following T cell activation in response to tBHQ as compared to PBMCs. tBHQ treatment increased production of the Th2 cytokines interleukin‐5, interleukin‐4, and interleukin‐13 at 120h by CD4 T cells after activation, and upon restimulation of these cells with anti‐CD3/anti‐CD28, indicating a potential effect on differentiation as well. Overall, these data indicate that Nrf2 inhibits primary human CD4 T cell activation, and potentially shifts differentiation towards a Th2 phenotype.

Support or Funding Information

(Supported by NIH grants ES018885 and ES007255).
Intracellular upstream signals activate histamine H1 receptor and interleukin‐9 gene expression, sensitive to pollinosis
Authors: Fukui Hiroyuki , Mizuguchi Hiroyuki , Kitamura Yoshiaki , Nemoto Hisao , Kashiwada Yoshiki , Takeda Noriaki
Published: November 2023

Number of pollinosis patients is increasing worldwidely. It is important for better alleviation to identify the sensitive genes of pollinosis. We observed that stimulation of histamine H1 receptor (H1R) induces H1R up‐regulation through the protein kinase C‐delta (PKC‐delta) activation pathway. Then we hypothesized that H1R up‐regulation worsens the symptoms of pollinosis due to the increase of receptor signaling. Correlation between nasal mucosal H1R mRNA level and symptom score was demonstrated in pollinosis patients, and treatment with antihistamine resulted the normalization of mRNA elevation and alleviation of symptoms. The result suggests that H1R gene functions as a sensitive gene for pollinosis.

Elevation of H1R mRNA was induced in nasal mucosa of allergic rhinitis model rats sensitized with toluene 2,4‐diisocyanete. The elevation was completely suppressed with antihistamine treatment. However, alleviation of symptoms was partial, suggesting the existence of the second gene, sensitive to pollinosis. Combination treatment of an antihistamine and Awa‐tea, a fermented tea produced in domestic area of Tokushima, Japan, was attempted. We observed that about 90% of symptom was alleviated. The result suggests that Awa‐tea contains a compound which suppresses gene expression of the second pollinosis sensitive gene.

Elevation of interleukin‐9 (IL‐9) mRNA level in nasal mucosa of allergic rhinitis model rats was suppressed with Awa‐tea. IL‐9 mRNA elevation was also observed in RBL‐2H3 cells, a mast cell line, through the activation of calcineurin (CN)/NFAT signaling. The elevation was suppressed with Awa‐tea, however, not with antihistamines.

An active compound was attempted to purify from Awa‐tea by monitoring suppressive activity on IL‐9 mRNA elevation, and pyrogallol was identified. Treatment with pyrogallol in combination with epinastine, an antihistamine, markedly alleviated nasal symptoms. Pyrrogallol did not inhibit protein phosphatase activity of CN. Pyrogallol suppressed ionomycin‐induced IL‐9 mRNA upregulation in RBL‐2H3 cells. Treatment with pyrogallol in combination with epinastine markedly alleviated nasal symptoms in allergic rhinitis model rats.

Above results suggest that combination therapy will be effective to alleviate allergic rhinitis by suppressing both PKC‐delta and CN/NFAT signaling pathways.
Intermittent Hypoxia and Hypercapnia Accelerate Aorta and Pulmonary Artery Atherosclerosis, partially Mediated via Trimethylamine‐Oxide (TMAO) in both ApoE−/− and Ldlr−/− Mice on High Fat Diet
Authors: Xue Jin , Zhou Dan , Poulsen Orit , Imamura Toshihiro , Hsiao Yu‐hsin , Malhotra Atul , Dorrestein Pieter , Knight Rob , Haddad Gabriel G
Published: November 2023

Obstructive sleep apnea (OSA) is a common disorder characterized by intermittent hypoxia and hypercapnia (IHC) during sleep and is often associated with increased inflammation, oxidative stress and dyslipidemia. Clinical studies have shown that OSA is an independent risk factor for atherosclerosis. To dissect the mechanisms involved, we compared the development of atherosclerotic lesions between two mouse models of atherosclerosis, i.e., the apolipoprotein E (ApoE) and the low density lipoprotein receptor (Ldlr) deficient mice, with or without IHC exposure. Ten‐week‐old ApoE−/− or Ldlr−/− mice were fed a high‐fat diet for 4 or 8 wks while being exposed to IHC 10 hr/day or normoxia 24 hr/day. En face lesions of the entire aorta, aortic arch and pulmonary artery (PA) were then examined. In addition, 3,3‐dimethyl‐1‐butanol (DMB), an inhibitor of microbial TMA production, was used to determine the contribution of microbes and TMAO in IHC‐induced atherosclerosis. The results showed that (1) 8‐week IHC exposure expedited high fat diet‐induced formation of atherosclerosis in both PA and aortic arch of ApoE−/− mice but only in PA of Ldlr−/− mice (ApoE−/− PA 8wks, IHC 35.4±1.9% vs Normoxia 8.0±2.8%, p<0.01); (2) the atherosclerotic lesions evolved faster and to a more severe extent in ApoE−/− mice as compared to those in Ldlr−/− mice (PA IHC 8wks, ApoE−/− 35.4±1.9% vs Ldlr−/− 8.2±1.5%, p<0.01); (3) DMB significantly attenuated IHC‐induced PA atherosclerosis but did not eliminate totally lesion formation induced by IHC (ApoE−/− PA 8wks, IHC DMB 25.6±2.6% either vs IHC 35.4±1.9%, p<0.01 or vs Normoxia 8.0±2.8%, p<0.01). Taken together, our findings suggest that IHC, a hallmark of OSA, accelerates the progression of atherosclerosis, in both vascular beds but especially in PA. This process is partly inhibited by a non‐lethal microbial TMA lyase inhibitor, suggesting the involvement of microbes and their metabolites TMA and TMAO, which may serve as potential therapeutic targets for the prevention and treatment of OSA‐induced atherosclerosis.
Investigating the Role of Arginine Methylation on Serine Phosphorylation in Histone H3
Authors: Mendiola Aron Judd Perez , Fuentes Daniel , Mendoza Mariel , Zurita‐Lopez Cecilia
Published: November 2023

We set out to investigate the effects of two PTMs, methylation and phosphorylation, with the goal of understanding the role of arginine methylation on neighboring serine phosphorylation in histone H3. Work by other labs has confirmed that arginine methylation and serine phosphorylation are involved in crosstalk within an AKT motif (RXRXXS/T) where the arginine residue is two residues away from the serine residue. Herein, we use the N‐terminal tail of histone H3 and focus on its arginine‐lysine‐serine (RKS) motif to determine whether a similar interaction is taking place. Serine‐10 hyper phosphorylation of histone H3 is a known biomarker for tumorigenesis, signaling the overexpression of proto‐oncogenes that lead to tumor formation, while arginine‐8 methylation of histone H3 is implicated in gene silencing. Preliminary results have determined that serine‐10 phosphorylation downregulates arginine‐8 methylation in histone H3 peptides. To further analyze the interaction, sequential in vitro methylation and phosphorylation reactions will be carried out using the respective protein arginine methyltransferase (PRMT) and kinase. Of the family of nine known human methyltransferases, emphasis is placed on PRMT1 and PRMT5, which are known to physiologically dimethylate histone H3. While there are several kinases that deposit the phosphate group on serine‐10, we will use mitogen and stress‐activated kinase 1 (MSK1) because it physiologically phosphorylates the serine‐10 residue and specifically targets RKS motif. Methodology for analyzing the methylation and phosphorylation interactions will be done through fluorography, western blot analysis, and thin layer chromatography analysis. Investigating the RKS motif will provide greater insights into protein function and can be a springboard for investigating these modifications in other proteins.

Support or Funding Information

NIH Grant #SC2GM118202
Interactive Role of TLR4 and TLR2 in Cardiac Functions During Stressful Conditions
Authors: Bagchi Ashim K. , Akolkar Gauri , Mandal Soma , Ayyappan Prathapan , Yang Xi , Singal Pawan K.
Published: November 2023

An appropriate balance between anti‐inflammatory interleukin‐10 (IL‐10) and pro‐inflammatory tumor necrosis factor‐α (TNF‐α) cytokines has been suggested for a normal functioning of the heart. It has been suggested that toll‐like receptor 4 (TLR4) promotes IL‐10‐mediated cardiac cell survival while another receptor, TLR2, from the same family is detrimental. We examined the interactive role of these two innate signaling molecules (TLR4 and TLR2) under stressful conditions including interleukin‐10 knockout (IL‐10−/−) mice, global ischemia/reperfusion (I/R) injury rat hearts and shRNA experimental models. Circulating and myocardial levels of TNF‐α as well as apoptosis and fibrosis were higher in IL‐10−/− hearts. Increase in TLR2 in IL‐10−/− hearts indicated its negative regulation by IL‐10. The ex‐vivo I/R also caused a marked upregulation of TLR2 and TNF‐α as well as apoptotic and fibrotic signals. However, 40 min reperfusion with IL‐10 in the I/R hearts, triggered an increase in TLR4 expression. Increase in interleukin‐1 receptor‐associated kinase‐M (IRAK‐M) and IRAK‐2 activity during I/R injury suggested their role in TLR2 signaling. Inhibition of TLR4 activity as a consequence of RNAi‐mediated suppression of myeloid differentiation gene 88 (MyD88) suggested a MyD88‐dependent activation of TLR4. Inclusion of IL‐10 during reperfusion, also significantly downregulated the expression of IRAK‐2, TRAIP and apoptotic signals, caspase 3 and Bax/Bclxl ratio. IL‐10 reduced the TNF‐α receptor‐associated increase in TRAIP/TRADD ‐induced apoptosis during ischemia injury which led to an increase in IL‐1β to mitigate TGF‐βRII‐mediated fibrosis. IL‐10 mitigation of these changes suggests that IL‐10 stimulation through TLR4 signaling, dissociates IRAK‐4 into IRAK‐1 instead of IRAK‐2 and may be an important therapeutic approach in restoring heart health from I/R injury.

Support or Funding Information

Supported by Canadian Institutes of Health Research and Research Manitoba.
Na2S, a Fast‐Releasing H2S Donor, Given as Suppository but not Parenteral GYY4137, a Slow‐Releasing H2S Donor, Lowers Blood Pressure in Rats
Authors: Ufnal Marcin , Tomasova Lenka , Drapala Adrian , Koszlewski Dominik , Ostaszewski Ryszard
Published: November 2023

Objectives

Although several H2S donors are available, their usefulness in research is questionable. Here, we evaluated hemodynamic effects of Na2S, a fast‐releasing H2S donor and GYY4137, a slow‐releasing H2S donor.

Methods

Hemodynamics were recorded in anesthetized Wistar Kyoto rats (WKY) and Spontaneously hypertensive rats (SHR) at baseline and after: 1) intraperitoneal (i.p.) infusion of either a vehicle (20% DMSO) or GYY4137 or Na2S; 2) intravenous (i.v.) infusion of either the vehicle or GYY4137 or Na2S; 3) per rectal administration of vehicle suppositories or suppositories with Na2S. Stability of GYY4137 in buffers and in rat plasma was evaluated with NMR.

Results

The vehicle and GYY4137 given i.p. and i.v. did not affect mean arterial blood pressure (MABP), whereas Na2S produced a significant, but transient (2–5 min) decrease in MABP. Administration of Na2S suppositories produced a dose dependent hypotensive response that lasted 60–90 min in WKY and SHR. In the buffers at pH of 7.4 and 5.5 and in rat plasma no reaction of GYY4137 was found during 18 hours of observation. In contrast, rapid decomposition of GYY4137 occurred in buffers at pH 2.0.

Conclusions

Na2S given in suppositories exerts a prolonged hypotensive effect in rats. GYY4137 does not affect MABP in rats, which seems to result from its high stability at physiological pH. Suppository formulation of fast‐releasing H2S donors may be useful in basic research as long as a reference slow‐releasing H2S donor is not available.

Support or Funding Information

This study was supported by The National Science Centre, Poland grant No. 2016/22/E/NZ5/00647
Multi‐scale systems analysis of blood pressure control in the spontaneously hypertensive rat
Authors: Beard Daniel Andrew , Bukowy John D. , Carlson Brian E. , Michele Daniel E. , Whitesall Steven E. , Van den Bergh Francoise
Published: November 2023

Hypertension and hypertensive disease are complex whole body syndromes that are unlikely to be effectively understood in terms of single‐cause/single‐effect relationships. One of the central barriers to understanding systems interactions in blood pressure control and hypertension has been an inability to formulate system‐level hypotheses in ways that yield experimentally testable predictions. To address this need, we have employed a modeling and simulation approach to analyze and interpret molecular, cellular, tissue, organ, and organ‐system data through developing multi‐scale models of components of the cardiovascular system responsible for processes ranging from beat‐to‐beat cardiovascular dynamics to long‐term regulation of blood volume. Using these models we have analyzed data from a battery of experiments to probe cardiovascular dynamics in the Spontaneously Hypertensive Rat (SHR) strain versus Wistar Kyoto (WKY) control strain. Integrating these model components together yields a cardiovascular system model that is able to represent the SHR and WKY strains, with differences identified primarily from in vivo measurements of cardiovascular mechanics (vascular pressures, volumes, flows), and neurohumoral variables (autonomic tone reflected by heart rate, endocrine factors) in response to blood volume perturbations. The identified model was used to test and represent the hypothesis that vascular mechanical remodeling represents a root cause of hypertension. Our results indicate that the mechanogenic mechanism is sufficient to explain the development of hypertension in the SHR rat.
Hypoxic Preconditioning Protects Rat Hearts Against Ischemia/Reperfusion Injury via H2S/TRPA1 Pathway
Authors: Ma Ming‐Chieh , Huang Ho‐Shiang , Chen Yih‐Sharng
Published: November 2023

Hypoxic preconditioning (HPC) protects rat hearts against ischemia/reperfusion (IR) injury. The Ca2+‐permeable transient receptor potential ankyrin 1 (TRPA1) is present in cardiac tissue and plays an important on blood pressure regulation. However, the role of TRPA1 in HPC‐mediated cardioprotection remains unknown. TRPA1 can be activated by a novel gasotransmitter hydrogen sulfide (H2S), which is synthesized endogenously by cystathionine β‐synthase (CBS) and cystathionine γ‐lyase (CSE). Here we examined whether HPC protects the myocardium against IR via the H2S/TRPA1 pathway. Compared to hearts of rats kept in room air, the hearts of rats kept in air preconditioned with 10% oxygen for 4 weeks had better post‐ischemic recovery and less tissue damage when subjected to 30‐min stop of perfusion and 4‐h reflow in a Langendorff apparatus. HC030031, a selective TRPA1 blocker, administered 5 min before reperfusion markedly attenuated the effects of HPC, confirming TRPA1 is a downstream effector in HPC‐mediated cardioprotection. Additionally, HPC markedly increased calcitonin gene‐related peptide (CGRP) and substance P (SP) release in coronary perfusate, which could be reversed by TRPA1 blockade, but not CGRP receptor or SP receptor inhibition. HPC resulted in the upregulation of CBS and CSE, and H2S levels in coronary perfusate, and prevented IR‐induced H2S reduction. Blockade of CBS and CSE by aminooxyacetic acid (AOAA) abrogated the effects of HPC and lowered release of H2S, CGRP, and SP. Mimicking the effects of HPC by given Na2S or NaHS to the AOAA‐treated hearts enhanced cardiac recovery and neuropeptide release during reperfusion. Our results demonstrate that HPC reduces heart susceptibly to IR via the H2S/TRPA1/neuropeptide pathway, as shown by increased enzyme responsible for H2S formation in HPC hearts.

Support or Funding Information

This study was supported by the Ministry of Science and Technology R.O.C. (MOST105‐2314‐B‐030‐009).
Imaging cardiac voltage gated sodium channels using the novel F‐18 radiotracer radiocaine
Authors: Schoenberger Matthias , Hooker Jacob , Strebl Martin , Schroeder Al , McKinsey Timothy , Ambardekar Amrut
Published: November 2023

Myocardial contraction is controlled by an electro‐mechanical coupling and cardiac action potentials that propagate across the heart. These action potentials are initiated by NaV1.5 voltage gated sodium channels. The role of cardiac NaV1.5 in cardiac disease is reasonably well understood in animal and in vitro models. However, linking changes in NaV1.5 expression levels to human disease in vivo has not yet been possible. “Radiocaine”, an F‐18 radiotracer for positron emission tomography (PET), is the first NaV imaging agent in the heart. Explants from healthy and failing human hearts were compared using radiocaine autoradiography to determine that the failing heart had ~30% lower NaV1.5 levels ‐ the first evidence of changes in cardiac NaV expression in humans as a function of disease. Paving the way for translational imaging, radiocaine proved to exhibit high in vivo specific binding to the myocardium of rats and non‐human primates. We envision that NaV1.5 measurements using PET imaging may serve not only as a powerful research tool, but also as a novel diagnostic to stratify arrhythmia risk and assess for progression of heart failure in patients with a broad spectrum of cardiovascular diseases.

Support or Funding Information

MS acknowledges experimental support by PD Dr. Heike Endepols, Dr. Mike Placzek. Work was supported by a MSCA‐IF‐GF grant to MS (INSCAPE 656644) and by the Phyllis and Jerome Lyle Rappaport MGH Research Scholar award (JMH). TAM was supported by the NIH (HL116848, HL127240 and AG043822) and the American Heart Association (16SFRN31400013). AVA is supported by a Scientist Development Grant from the American Heart Association, the Boettcher Foundation's Webb‐Waring Biomedical Research Program, and in part by NIH/NCATS Colorado CTSA Grant Number UL1 TR001082. Contents are the authors' sole responsibility and do not necessarily represent official NIH views. This work was in part supported by access to equipment purchased using NIH shared instrument grants S10RR017208 and S10RR023452.
Effect of All‐Extremity Exercise Training on Left Ventricular Function Assessed by Speckle Tracking Echocardiography in Healthy Older Adults
Authors: Hwang Chueh‐Lung , Petersen John W , Mackay Karen , Yoo Jeung‐Ki , Kim Han‐Kyul , Hwang Moon‐Hyon , Handberg Eileen M , Szady Anita D , Christou Demetra D
Published: November 2023

Aging is associated with cardiac remodeling and alterations in cardiac function. Recent advances in two‐dimensional echocardiography have allowed for detailed assessment of left ventricular (LV) function using speckle tracking echocardiography (STE). The purpose of this study was to compare the effect of all‐extremity high‐intensity interval training (HIIT) versus moderate intensity continuous training (MICT) on STE‐determined measures of LV systolic and diastolic function in healthy older adults. A total of 50 sedentary older adults (age 65±1.0 years, 17 men) were randomly assigned to HIIT (n=17), MICT (n=17) or non‐exercise controls (CONT, n=16). HIIT (4×4 minutes at 90% of peak heart rate) and isocaloric MICT (70% of peak heart rate) were performed on an all‐extremity non‐weight‐bearing ergometer, 4 times per week, for 8 weeks under supervision. Resting LV global longitudinal strain (S) and strain rate (SR) were measured using two‐dimensional STE of 4‐, 2‐, and 3‐chamber views to assess systolic function (Ssys and SRsys) and diastolic function (early and late SRdia). Thirty‐nine subjects completed the intervention. LV Ssys (HIIT: −19.8±0.9 vs. −20.3±0.9; MICT: −18.2±1.0 vs. −19.7±0.9; CONT: −20.6±0.9 vs. −20.1±0.7), SRsys (HIIT: −1.12±0.06 vs. −1.14±0.05; MICT: −1.06±0.06 vs. −1.20±0.06; CONT: −1.24±0.05 vs. −1.20±0.04), early SRdia (HIIT: 1.13±0.06 vs. 1.19±0.07; MICT: 1.07±0.10 vs. 1.15±0.09; CONT: 1.28±0.07 vs. 1.28±0.10), and late SRdia (HIIT: 0.68±0.06 vs. 0.69±0.05; MICT: 0.56±0.05 vs. 0.71±0.06; CONT: 0.68±0.05 vs. 0.66±0.05) did not change in response to the intervention (P>0.2). In conclusion, STE‐determined LV systolic and diastolic function were unaffected by 8 weeks of all‐extremity exercise training in healthy older adults.

Support or Funding Information

This work was supported by NIH AG 050203 to DDC.
Appearance of Local Abnormalities of Left Ventricular Contraction on Top of Global Diastolic Dysfunction in the Hypertensive mRen2 Rat
Authors: Csipo Tamas , Fulop Gabor Aron , Bodi Beata , Fagyas Miklos , Priksz Daniel , Juhasz Bela , Beke Livia , Hendrik Zoltan , Mehes Gabor , Olah Attila , Radovits Tamas , Merkely Bela , Granzier Henk , Papp Zoltan , Judit Barta , Toth Attila , Kovacs Arpad
Published: November 2023

Hypertension is a major co‐morbidity of diastolic dysfunction. Appearance of systolic dysfunction on top of diastolic dysfunction further increases the risk of cardiac failure and mortality. We hypothesized that a hypertensive rat model with diastolic dysfunction can give insight into the pathomechanism of progressing human heart failure.

To this end we compared 15 weeks old male rats (n=9‐9) with hypertension due to a renin transgene (mRen2, 228.9±4.9 mmHg; P<0.05) with geno‐/phentoype controls (SD, 121.3±4.4 mmHg).

Invasive parameters of left ventricular (LV) active relaxation (Tau, mRen2: 16.9±0.8 ms vs. SD: 12.6±0.6 ms; P<0.05) and passive stiffness (slope of EDPVR, mRen2: 0.050±0.004 mmHg/μL vs. SD:0.040±0.002 mmHg/μL; P<0.05) confirmed diastolic dysfunction with preserved ejection fraction (EF, mRen2: 52.5±0.7% vs. SD:53.3±0.7%) in the mRen2 animals.

In contrast, regional echocardiography (n=8‐8) and histology (n=5‐5) showed worse Tei index (0.78±0.04; P<0.05) and relatively higher wall thickness (0.74±0.06; P<0.05) of the LV free wall in mRen2 rats than those of the interventricular septum (0.68±0.02 and 0.54±0.03, respectively).

At the cellular level, both lower Ca2+ sensitivity (pCa50) and higher troponin I (cTnI) phosphorylation of LV cardiomyocytes from the free wall and the septum (n=mean of 3‐3/4‐4 rats) suggest abnormal systolic function in the mRen2 group as compared to the control. LV heterogeneity of mRen2 rats is further supported by lower cardiomyocyte maximal Ca2+‐activated force (21.2±1.4 kN/m2), higher Ca2+ sensitivity (5.85±0.01) and phosphorylation of cTnI at Thr‐144 (1.62±0.19; P<0.05) in the free wall than those in the septum (30.0±0.6 kN/m2, 5.76±0.02 and 1.00±0.15, respectively).

The local abnormalities of LV contraction on top of global diastolic dysfunction may provide a pathomechanism for the clinical progression with increasing mortality seen in human.
A Histological Comparison of Ear Skin Regeneration in Acomys and Mus
Authors: Sandoval Aaron , Brant Jason , Maden Malcolm
Published: November 2023

Regeneration has been studied almost exclusively in lower invertebrates as most mammals are only able to regenerate fetal tissue. The African spiny mouse (Acomys) represents the first time advanced regeneration has been observed in an adult mammal. Acomys has evolved a defense mechanism which involves fragile skin that tears easily when caught by a predator, allowing the mouse to escape. Subsequently, the mouse regenerates extensive parts of its body. The regenerative capabilities of Acomys are being studied by comparing it to a normal mouse (Mus). In order to compare the progression of ear regeneration in Acomys and Mus, ears of both species were wounded using a four‐millimeter punch to remove the epidermal and dermal tissue layers, revealing the underlying cartilage. The healing ears were harvested at 2, 4, 7, 14, 21, and 30 day time points. The ears were subsequently embedded in wax, mounted on slides, and trichrome stained to differentiate between erythrocytes, muscle and collagen. Microscopic analysis revealed that although the cartilaginous layer eventually degenerated in both species, extensive degeneration was present much earlier in Mus. Furthermore, Acomys was able to regenerate its cartilage and hair follicles, whereas Mus was only able to regenerate a disorganized, nonfunctioning mass of collagen. Significant scarring was evident in Mus, while no scarring was observed in Acomys. The results of further study of Acomys could prove integral in gaining a comprehensive understanding of the regenerative process. Findings could ultimately improve the entire healthcare field by allowing for the regeneration of human tissue.
Angiotensin II‐mediated Proliferation of Neonatal Cardiac Fibroblasts and Role of Mitochondrial Ca2+ Uptake and Superoxide Generation
Authors: Mishra Jyotsna , Fu Deming , Jhun Bong Sook , Sheu Shey‐Shing , O‐Uchi Jin
Published: November 2023

Cardiac fibroblasts (CFs) are the major cell type in addition to cardiomyocytes in the mammalian heart, and they play a vital role in regulating normal cardiac function and adaptive cardiac remodeling during adverse clinical conditions, including fibrosis. Recent studies have revealed Angiotensin II (AngII) as a one of the principal effectors for promoting fibroblast differentiation and cardiac remodeling in the failing heart. In addition, evidences have shown that the downstream effects of AngII are mediated by elevated levels of reactive oxygen species (ROS) which have also been implicated in CF proliferation. However, the detailed signaling mechanism of ROS generation by AngII signaling in the CFs still remains unclear. We previously demonstrated in cardiac myoblast H9C2 cells that the main Ca2+ uptake mechanism, the mitochondrial Ca2+ uniporter (MCU)‐mediated mitochondrial Ca2+ overload, induces mitochondrial superoxide (mtSO) generation. Therefore, we hypothesize that Ang‐II mediated stimulation may increase mitochondrial Ca2+‐induced mtSO generation, which may further activate ROS‐dependent proliferation signaling in CFs. Using mitochondria‐targeted Ca2+ biosensor in neonatal rat CFs (NCF), we first established that AngII (≥1 μM) stimulation significantly increases the mitochondrial Ca2+ uptake in response to cytosolic Ca2+ elevation induced by the Ca2+ release from the endoplasmic reticulum (ER). Further, we also found that AngII stimulation leads to increase in the mtSO levels. We next confirmed that proliferative pathways, including ERK1/2 and p38 AT‐II were also activated in a time‐dependent manner on AngII stimulation in CFs, while the effect was abolished by pretreatment with mitochondria‐targeted antioxidant, Mito‐tempo without affecting the profiles of AngII ‐induced ER Ca2+ release and mitochondrial Ca2+ uptake. Finally, genetic inhibition of MCU function by overexpressing dominant‐negative MCU also blocked AngII‐mediated activation of proliferative pathways in CFs. In conclusion, these data suggest that MCU‐mediated mitochondrial Ca2+ accumulation induces mtSO generation and this ROS generation serves as an important regulator for the proliferative pathway in CFs under Ang II stimulation.

Support or Funding Information

• 14BGIA18830032 and 16SDG27260248 from AHA to J.O‐U.

• Medical Research Grant (#H1403) from W.W. Smith Foundation to J.O.‐U.

• NIH grant (R01 HL093671) to S.‐S.S
Green fluorescence from non‐aequorea hydrozoa Gonionemus vertens (Clinging Jellyfish)
Authors: Petroff Elena , Snitsarev Vladislav , Lopez Rafael , Bologna Paul A.X. , Gaynor John J.
Published: November 2023

We have examined the green fluorescence of the Clinging Jellyfish, Gonionemus vertens (GV), a species that first appeared in New Jersey coastal estuaries in June, 2016. Intact specimens (0.30–0.35 g wet weight) were homogenized in 3 mL of pure water using a Kontes Glass homogenizer (Co 23; Vineland, NJ) with 10 rotational strokes and centrifuged at 16,100 × g for 10 min at 4°C (Eppendorf 5415D microcentrifuge). Two mL of the supernatant was placed in a standard 1 cm methacrylate cuvette and fluorescence was measured with a Hitachi F‐7000 spectrofluorometer at 25°C. In vitro excitation and emission maxima were λExMax = 501.4±0.2 (nm) (n=4) and λEmMax = 512.9±0.2 (nm) (n=3), respectively for the 4 individuals measured.

Adult medusa of GV are relatively small and fit in a standard 1 cm spectroscopic cuvette, so we measured in vivo fluorescence of the intact individuals in sterile artificial seawater (20 ppm). When first placed in the cuvette, actively swimming GV cannot be measured; but after some time, individuals use their tentacles to cling to the inner cuvette wall and remain stationary permitting measurements. Multiple spectral scans from 450 nm to 510 nm (excitation scan) or from 510 to 570 nm (emission scan) at 240 nm/min (15 seconds per scan) were performed. In five individuals successfully measured without movements, excitation and emission maxima were λExMax = 503.9±0.4 (nm) (n=5) and λEmMax = 515.4±0.2 (nm) (n=5), respectively. t‐tests between in vitro and in vivo measurements showed a significant difference between both excitation maxima (p<0.001) and emission maxima (p<0.0001), indicating that the green fluorescence of GV is sensitive to environmental parameters.

A search of the literature of green fluorescent proteins suggests that the particular excitation/emission pattern we have recorded in GV is unique and represents the shortest Stokes shift seen in nature. We plan to continue our studies on the green fluorescence of GV with a goal of isolating both the gene and protein responsible for this fluorescence through transcriptome analysis.

Support or Funding Information

Sokol grant to JG, VS, and EP
Neuroimmune communication between the circadian clock and tumor development in mice
Authors: Aiello Ignacio , Roman Fernanda , Fedele Malena Mul , Cerliani Belen , Richard Silvina , Chiesa Juan Jose , Golombek Diego Andrés , Paladino Natalia
Published: November 2023

Daily environmental changes on Earth have imposed a selective pressure for life driving the development of circadian clock mechanisms that generate rhythms in physiological and behavioral variables, and are capable of entraining to external cues. In mammals, the clock resides in the hypothalamic suprachiasmatic nuclei (SCN), and the principal signal that adjusts its activity is the light‐dark (LD) cycle. Bidirectional interactions between the immune and the circadian systems have been under intensive study in recent years. Particularly, it has become very important the role of the circadian clock in tumor progression. According to the World Health Organization, work schedules that imply a desynchronization between the circadian system and the environmental and social factors, such as shift work, represent a risk factor for the development of cancer. Moreover, it's known that tumor growth can be regulated by genes that are part of the molecular circadian clock.

Our purpose is to determine the effect of circadian desynchronization induced by chronic jet‐lag (CJL) in tumor development, focusing in the role of the immune system. We injected subcutaneously B16 cells (murine melanoma) in C57bl/6 mice housed under CJL (6 h advances every 2 days) or normal light (LD 12:12) conditions. Our results show that the tumor growth rate was significantly higher, while latency was lower under CJL condition. However, early tumor angiogenesis was not affected. In addition, tumor and liver expression of clock genes Per1–2 were decreased under CJL condition, but levels of the clock gene Bmal1 were similar under both conditions. We have also studied the proinflammatory response in the tumors of these animals at two different time points, and we found that the levels of cytokines TNF‐α, IL‐6 and IL–1β were induced during the day in animals living in normal light conditions, while the expression was similar at both time points in animals subjected to circadian desynchronization. Additionally, the spleen percentage of mature antigen‐presenting cells was decreased under CJL schedule.

For the other hand, animals carrying tumors showed a decrease in the percentage of nocturnal activity and in the strength of their locotomor activity rhythms in LD conditions. We have also analyzed the mRNA levels of the chemokine Ccl2/MCP‐1 (Monocyte Chemotactic Protein‐1) and TNF‐α in the SCN of animals subjected to normal light conditions. The first one seems to increase in the SCN of animals with the tumor, while the other one was undetectable.

In conclusion, here we show that the bidirectional interaction between the immune and circadian systems can be mediated by cytokines/chemokines in a murine model tumor development. We have also confirmed that circadian desynchronization can affect the development of a tumor and that the inflammatory molecules present in the tumor are over circadian control.
Hibernation: Insights from Comparative Genomics of Zapus Species
Authors: Israelsen William J. , McNulty Alyssa D. , Cong Qian
Published: November 2023

Hibernating mammals employ unique metabolic strategies to survive harsh conditions. They become obese prior to the onset of winter, then conserve their stored energy during a long winter fast by slowing metabolism and becoming deeply hypothermic. Hibernation phenotypes may provide insights into control of metabolism and have applications in medicine, but little is known about the molecular mechanisms or genetic underpinnings of hibernation. We have begun using the meadow jumping mouse (Zapus hudsonius) as a laboratory hibernation model. We sequenced, de novo assembled, and annotated a meadow jumping mouse genome. We further generated whole genome sequences for six additional meadow jumping mice and seven western jumping mice (Zapus princeps), a closely related hibernator. Z. hudsonius uses photoperiod as the cue to initiate fall fattening, while Z. princeps ignores photoperiod and prepares for hibernation based on food availability. Preliminary comparative genomic analysis reveals a subset of ~1700 proteins that are significantly diverged between the species. These diverged proteins are enriched for genes involved in pheromone sensing and reproduction, immunity, and metabolism. Analysis of individual genes yields insights into speciation and may reveal candidate genes that explain the phenotypic differences between the two species.

Support or Funding Information

WJI acknowledges support from the Sara and Frank McKnight Fund for Biochemical Research and an NIH Early Independence Award (DP5OD021365).
Effects of amaranth (Amaranthus esculantus) and canola (Brassica napus) oil supplementation on oxidative metabolism of neutrophils in patients with obesity
Authors: Dominika Kanikowska , Alina Kanikowska , Rafał Rutkowski , Małgorzata Włochal , Marian Grzymisławski , Maki Sato , Andrzej Bręborowicz , Janusz Witowski
Published: November 2023

Reactive oxygen species are postulated to be involved in systemic inflammation and oxidative stress in patients with obesity. Activated polymorphonuclear neutrophils (PMNs) generate high amounts of reactive oxygen species. The aim of the study is to investigate whether amaranth oil (Amaranthus esculantus) and canola (Brassica napus) oil supplementation impacts on oxidative metabolism in patients with obesity. The hypothesis is that, owing to its lipid‐lowering and antioxidant properties, amaranth oil and canola oil will protect against oxidative stress. In the study, the effects of amaranth and canola oils on oxidative metabolism and apoptosis of neutrophils from obese individuals has been investigated. We tested 19 obese patients (aged 48.3±16 years,6 women and 13 men, BMI = 41.1 ±7.8 kg/m2, (mean ±SD)). The study adhered to basic Principles of the Declaration of Helsinki. Neutrophils were isolated and oxidant production, in response to phorbol 12‐myristate 13‐acetate, was characterized by using luminol dependent chemiluminescence (CL) and flow cytometric dichlorofluorescin oxidation assay. Neutrophils from obese individuals before treatment had a significantly higher H2O2 production and CL response compared to obese after 3 weeks of both oils supplementation (p < 0.05). This amaranth oil and canola oil exert beneficial effects in cells and may, therefore, be useful in the treatment of obesity disorders.

Support or Funding Information

none