bluuebunny/crossref_metadata_2025_split huggingface dataset:
LIM kinase inhibition reduces endothelial cell cortical stiffness in diabetic arteries
Authors: Power Gavin , Gonzalez-Vallejo Juan , Ramirez-Perez Francisco , Lateef Olubodun , Manrique-Acevedo Camila , Martinez-Lemus Luis , Padilla Jaume 
Published: May 2023

Type 2 diabetes is associated with endothelial stiffening, which precedes the development of increased arterial stiffness. The role of endothelial cell stiffening in the pathogenesis of vascular disease is increasingly recognized. Indeed, evidence exists that cellular pathways that result in polymerization of endothelial cell actin filaments (F-actin) and increased cellular stiffness impair the formation of nitric oxide, a potent vasodilator and anti-inflammatory molecule. We and others have shown that, in vascular smooth muscle cells, LIM kinase (LIMK) is a critical regulator of actin dynamics and cellular stiffness via the phosphorylation and consequent inactivation of the F-actin severing molecule, cofilin. However, whether LIMK regulates actin polymerization and stiffness in endothelial cells remains unknown. Here we report that pharmacological inhibition of LIMK with LIMKi3 in cultured human endothelial cells for 18 hours decreases cofilin phosphorylation and cortical stiffness, as assessed by atomic force microscopy. These endothelial softening effects are accompanied with a reduction in F-actin content. Reduced F-actin in endothelial cells is also recapitulated with S3, a cofilin mimetic. Furthermore, we show that the aortic endothelium of diabetic ( i.e., db/db) male mice is stiffer than that of wild-type counterparts, and that stiffer diabetic arteries also exhibit a tendency for increased p-cofilin. Notably, treatment of diabetic aortas with LIMKi3 for 18 hours also reduces endothelial cortical stiffness. Collectively, our findings demonstrate that LIMK inhibition reduces endothelial cortical stiffness in naïve cultured endothelial cells and in diabetic arteries ex vivo. These findings support the hypothesis that LIMK may serve as a potential target for reversing endothelial stiffening in type 2 diabetes. Funding: R01HL151384, R01HL153264, and R01HL137769 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Oral Carnosine Supplementation Restores Insulin Sensitivity but not Glucose Tolerance in Mice With Glutathione Peroxidase-4 Deficiency
Authors: Berdaweel Islam , Mahoney Jo , Alowaisi Amany , Berns Kaitlyn , Anderson Ethan 
Published: May 2023

Obesity presents in about 90% of the diabetic patients, a risk factor that makes them more susceptible to cardiometabolic disorders, particularly insulin resistance and ultimately T2 diabetes. Increased oxidative stress and specifically lipid peroxidation are known to be major pathogenic factors driving these disorders in obesity. We recently reported that diabetic patients were found to have significantly lower level of glutathione peroxidase 4 (GPx4)- an antioxidant selenoenzyme which neutralizes lipid peroxides. Carnosine, a histidine-containing dipeptide which is highly concentrated in muscle and brain, has shown a remarkable therapeutic potential for preventing diabetes and its complications when administered orally in experimental and clinical studies of obesity. The extent to which carnosine can mitigate these disorders in the presence of GPx4 deficiency is not known however, and this is critical to determining its efficacy in diabetes patients. To address this question, wild-type (WT) and GPx4+/− male were randomly assigned to either standard chow (CNTL) or high fat high sucrose diet (HFHS) for 16 weeks. Seven weeks after starting the diet, half of the HFHS diet groups received 80 mM carnosine oral carnosine supplementation in their drinking water. Glucose and insulin tolerance tests performed 8 weeks after starting the diet revealed that carnosine significantly improved glucose tolerance in obese WT (P<0.05) but not obese GPx4+/− mice. Interestingly, carnosine treatment did significantly improve insulin sensitivity in the GPx4+/− mice, and to a lesser extent the WT mice. At the molecular level, we observed that HFHS diet up-regulated GPx4 protein in cardiac tissue from WT but not GPx4+/− mice, and protein oxidation was significantly reduced with carnosine treatment in both genotypes. Collectively, these findings indicate that carnosine supplementation could potentially mitigate insulin resistance even in patients with compromised antioxidant capacity (e.g., diabetes). Further analysis is in progress to evaluate the effectiveness of carnosine supplementation on additional cardiometabolic complications that are associated with diet induced obesity. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Epithelial heme oxygenase-1 promotes colitis associated tumorigenesis through regulation of tumor epithelial differentiation and survival
Authors: Callahan Rosemary , Onyiah Joseph , Bhagavatula Geetha , Schaefer Rachel , Staley Alyse , Murphy Emily , Zhou Liheng , Orlicky David , Atif Shaikh , Cartwright Ian , Hall Caroline , Colgan Sean 
Published: May 2023

Damage from chronic inflammation in the setting of inflammatory bowel disease has been linked to increased rates of small and large bowel cancer. Colonic inflammation is often associated with hemorrhage resulting in release of heme from damaged erythrocytes. Heme oxygenase-1 (HO-1) protects against oxidative DNA damage from heme. We hypothesized that deletion of intestinal epithelial cell (IEC) HO-1 would increase colitis associated tumorigenesis. Inflammation associated hemorrhage correlated with mucosal Hmox1 expression in a murine model of colitis, but conditional deletion of IEC Hmox1 (Hmox1ΔIEC) did not influence inflammatory outcomes. However, in a colitis associated cancer model, we unexpectedly observed decreased number and size of colonic tumors in Hmox1ΔIEC mice compared to controls. Single cell RNA sequencing revealed a similar tumor leukocyte profile, but the tumor epithelial population displayed increased colonocyte differentiation in Hmox1ΔIEC mouse tumors relative to stem/secretory epithelial populations. Deficiency of HO-1 also lead to increased cell death and cell cycle arrest in murine colonoids and colon cancer cells, which was exacerbated by heme. Findings define a previously unreported role for epithelial HO-1 in supporting tumorigenesis in the colitic microenvironment. US Department of Veterans Affairs This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
The Role of Mitochondrial ABCB8 in Exercise-Induced Protection Against DOX Cardiotoxicity
Authors: Doerr Vivian , Montalvo Ryan , Jacintho Moritz Cesar , Boeno Franccesco , Dowllah Imtiaz , Smuder Ashley 
Published: May 2023

Doxorubicin (DOX) is a highly effective chemotherapeutic used in cancer treatment. However, its use is associated with the development of cardiac dysfunction, which can negatively affect patient outcomes and reduce overall survival. While no clinical countermeasure exists to combat DOX cardiotoxicity, endurance exercise preconditioning is sufficient to preserve cardiac function following acute DOX exposure. In this regard, it is established that DOX cardiotoxicity is associated with increased mitochondrial reactive oxygen species (ROS) production and iron accumulation and that endurance exercise training stimulates an increase in the expression of the mitochondria-localized ATP-binding cassette subfamily B member 8 (ABCB8) transporter. Overexpression of ABCB8 has been shown to prevent DOX cardiac dysfunction. Therefore, we tested the hypothesis that increased levels of ABCB8 are required for exercise-induced cardioprotection against DOX toxicity. Cause and effect were determined by preventing the exercise-induced increase in ABCB8 expression using an antisense oligonucleotide (AO) designed to specifically target ABCB8. Young adult female Sprague-Dawley rats remained sedentary (SED) or were treadmill exercise trained (EX) for 2 weeks (70% VO2MAX, 1hr/day, 10 days). Immediately following each exercise bout or at identical time points, rats received either AO (10mg/kg i.p.) or saline. Rats then received either a bolus dose of DOX (20mg/kg i.p.) or saline (SAL) treatment 24 hours after the last exercise bout. Dependent measures were evaluated 48 hours after DOX or saline administration. Body weight change following DOX/SAL treatment showed significant weight loss in all DOX-treated groups, with the greatest weight change occurring in the EX-AO-DOX rats. Additionally, EX-AO-DOX treatment prevented the rescue of heart weight/tibia length seen in the EX-DOX group. Exercise capacity was assessed via a graded exercise tolerance test and revealed a significant reduction in running time in both the SED-DOX and EX-AO-DOX groups compared to SED-SAL. Interestingly, mitochondrial ROS emission was not elevated in the EX-AO-DOX treatment group compared to SED-SAL. Collectively, these results reveal that exercise-induced ABCB8 expression contributes to the protective effect of exercise against DOX cardiotoxicity. NIH R01 HL146443, NIH R01 HL144858, T32 HD043730 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Generating interactive visual maps of anatomical connectivity from SPARC connectivity knowledge
Authors: Nickerson David , Brooks David , Balachandran Biruthuvan , Gillespie Tom , Imam Fahim , Grethe Jeffrey , Tappan Susan , de Bono Bernard , Martone Maryann , Hunter Peter 
Published: May 2023

The December 2022 release of the SPARC Portal ( https://sparc.science ) included the first significant update to the anatomical connectivity flatmaps since the portal first launched. These flatmaps provide an interactive and visual map for the display and exploration of the autonomic nervous system of Human, rat, mouse, pig, and cat ( https://sparc.science/maps ). In addition to improved anatomical ‘cartoons’ for all species and adding a female Human map, these maps, for the first time, automatically render the connectivity knowledge directly retrieved from the SPARC Connectivity Knowledge Base of the Autonomic Nervous System (SCKAN; https://sparc.science/resources/6eg3VpJbwQR4B84CjrvmyD ). SCKAN contains explicit knowledge about CNS-ANS-end organ circuitry derived from SPARC data and scientific literature, in a form that supports computational reasoning. Each flatmap consists of a manually drawn base layer for the species-specific anatomical cartoon and a layer of manually drawn tracts for the large nerves. By annotating these drawings with standard reference SPARC vocabularies consistent with SCKAN usage, software tools are then able to semantically map connectivity circuits retrieved from SCKAN to the visual representation on each species’ flatmap. Beyond the interactive visual rendering of the circuitry on the SPARC Portal, the semantic consistency between flatmaps and SCKAN powers further user interface components on the SPARC Portal to surface additional knowledge for each rendered connection. For example, comprehensive links to the literature and/or data supporting a connectivity statement can be retrieved from SCKAN and provided to Portal users. Now that tools are in place to support this automated workflow to generate the flatmaps from SCKAN knowledge, we are continuing to improve the SPARC Portal to visualise new knowledge as it becomes available. NIH Common Fund, NIH Office of the Director, Awards OT3OD025349 and OT2 OD030541 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Suppression of hematologic and neurological expression-1 promotes tumor immunity by regulating the GSKβ-PD-L1 Axis
Authors: Kim Soo Mi 
Published: May 2023

Despite various efforts have been made to prolong the liver patients’ survival rates, liver cancer is still one of the malignancies with a high mortality rate in worldwide. The clinical efficacy of anti-PD-1/PD-L1 treatments has been investigated associated with PD-L1 levels, but, the relationship between hematologic and neurological expression 1(HN1) and immunosuppressive molecules PD1 and PD-L1 in liver cancer is not clearly understood. Here, we reported for the first time that HN1 acts as a novel negative regulator of PDL1, whose abnormal expression plays a crucial role in cancer immune evasion in hepatocellular carcinoma. This study establishes a key molecular link between targeted therapy and immune surveillance and identifies that combination therapy with HN1 inhibition and an anti-PD-1 antibody has much better antitumor efficacy than either monotherapy in hepatocellular carcinoma. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Potential roles of BNIP3 mediated placental mitophagy in the development of gestational diabetes mellitus
Authors: Gao Haijun , Wu Bin , Gao Albert , Mitchell Rochanda , McDonald Vanessa 
Published: May 2023

To date, the root cause of Gestational diabetes mellitus (GDM) remains unclear. A handful of studies found that mitochondrial defects occur in the GDM placenta, including excess accumulation of destroyed mitochondria, reduced ATP and enhanced ROS production, which indicates that mitophagy may be impaired in the GDM placenta. In this study, we hypothesized that placental mitophagy mediated by BNIP3 plays a critical role in the development of GDM. In Study One, to investigate whether mitophagy is impaired in the GDM placenta, we compared the protein abundance of mitophagy markers (LC3II, SQSTM1) or major pathway mediators (PRKN, BNIP3, FUNDC1) in mitochondrial factions of placental tissues collected from women with normal glucose tolerance (NGT) or GDM at term pregnancy, following Western blotting analyses. In Study Two, we knocked down BNIP3 expression in human trophoblast cell line BeWo using Lentivirus siRNA transfection, then mitochondrial ATP production was measured by Seahorse Mito Cell Stress Test. Study Three, the changes of transcriptome in response to BNIP3 knockdown (BKD) was investigated by RNA sequencing. All numerical parameters between the NGT and GDM groups, between the BKD and control (CT) cells were analyzed by ANOVA (n=4 or 3). The main findings include: 1) The protein abundance of LC3II, PRKN, BNIP3, FUNDC1 in the mitochondrial fractions of GDM placentas was reduced by 1.6-, 1.7-, 1.3-, and 1.5-fold, compared to that in NGT placentas respectively, while that of SQSTM1 was increased by 1.9-fold. 2) Mitochondrial ATP production at basal levels was reduced by 1.2-fold in BKD cells, compared to CT cells. Similarly, spare respiratory capacity, ATP-production coupled respiration, maximal respiration and non-mitochondrial oxygen consumption were reduced by 1.7-, 1.3-, 1.4- and 1.6-fold in BKD cells, respectively. 3) A total of 382 differentially expressed genes were identified in BKD cells, with 158 genes upregulated and 224 genes downregulated. Among these genes, expression of four mitochondria related genes TOMM6, MT-ATP6, NOL3 and IFI6, was reduced by 9.1-, 2.2-, 1.6-, and 1.5-fold in BKD cells, compared to CT cells, respectively, while the expression of MCUB was increased by 1.5-fold. These results suggest that BNIP3 and BNIP3 mediated mitophagy pathway may play an important role in mitochondrial homeostasis in human trophoblast cells, and the reduced BNIP3 expression in GDM placentas may lead to mitochondrial structural and functional defects, thus affecting placental functions and contributing to the development of GDM. NIH research grants R03HD095417, U54MD007597 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Human KCC2a Protein Expression and Phosphorylation is Altered by Elimination of the Second Cryo-EM Identified Chloride Binding Site
Authors: de los Heros Paola , Canto Aranza , Mercado Adriana 
Published: May 2023

Background: Potassium-chloride cotransporters (KCCs) are responsible for the symport of Cl- with K+ in an electroneutral manner. They play important roles in numerous crucial physiological processes, from cell volume regulation to transepithelial solute transport, intracellular ion homeostasis and regulation of GABA neurotransmission. Under isotonicity, except for KCC2, KCCs remain inhibited due to phosphorylation of residues located in cytoplasmic domains, whereas in response to hypotonic cell swelling, dephosphorylation of these residues stimulates transport activity. After several decades of studies, new breakthroughs with the cryo-EM structural determination of KCCs have deepened our understanding of their molecular basis and transport function, including architecture, ion binding sites, and coupled ion transport. In the present study, we mutated and analyzed the two chloride-binding sites in human KCC2a recently reported by cryo-EM to determine whether the phosphorylation status is modified upon their removal. Methods: Xenopus laevis oocytes and HEK-293 cells were either microinjected or transfected with human KCC2a wild type (WT) or chloride-binding site mutants, G136/V137/I138A (S1-Cl), and G436/I437/M438/Y592A (S2-Cl). KCC2a phosphorylation status in response to isotonic and hypotonic conditions was evaluated by immunoblot analysis of regulatory phosphorylation sites pThr926 and pThr1030. In parallel, phosphorylation status of SPAK/OSR1 activating regulatory site pSer373/pSer325 was analyzed under the same conditions. Results: Elimination of either chloride-binding site induced changes in total expression of KCC2a. Removal of S1-Cl slightly increased total KCC2a expression, whereas lack of S2-Cl showed a significant reduction of total protein in Xenopus laevis oocytes and HEK-293 cells. Phosphorylation of regulatory Thr1030 in S2-Cl mutant is significatively decreased on both biological models, even under isotonic inhibitory (phosphorylated) conditions. pThr1030 on S1-Cl mutant behaves similarly to WT KCC2a. Phosphorylation of regulatory SPAK kinase is not affected on both mutants. Conclusions: Our results suggest that while both sites affect KCC2a total expression, only the S2-Cl appears to be affecting KCC2a activating response. It appears S2-Cl mutation dysregulates SPAK signaling pathway, since the kinase osmotic response remains unchanged on presence of KCC2a chloride binding mutants. AM INCar 22-1345 and PdlH UNAM Papiit IN222320 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Vasoactive Intestinal Peptide Reverses The Impaired Airway Relaxation in Neonatal Rats Exposed to Hyperoxia
Authors: Sopi Ramadan , Reçica Shkëlzen , Kryeziu Islam , Basholli-Salihu Mimoza , Orqusha Nimet , Mladenov Mitko , Neziri Burim , Thaçi Qëndrim 
Published: May 2023

Neonatal hyperoxia increases contraction and decreases the relaxation of airway smooth muscle (ASM), and the balance between these processes in airways is critical for normal flow of air. Thus, it is important to search for an effective treatment that targets this balance to prevent the airway hyperreactivity. The non-adrenergic- noncholinergic inhibitory system mediators such as vasoactive intestinal peptide (VIP) play an important role in ASM tone. Therefore, we tested the hypothesis that exogenous VIP will reverse the impaired relaxation of tracheal smooth muscle exposed to neonatal hyperoxia. Tracheal cylinders were obtained from Wistar rat pups (P10) exposed to hyperoxia (≥ 95% O2) or room air for seven days. These cylinders were used to study relaxant responses of tracheal smooth muscle (TSM) evoked by electrical field relaxation (EFS) in precontracted tissue (carbachol, 10μM) in absence or presence of a single dose of VIP (50 nM). In addition, the dose-response relaxant effect of VIP (1nM – 1μM) was tested in absence or presence of a VPAC receptor antagonist; a protein kinase A (PKA) inhibitor (Rp-8-CPT-cAMPS); a phosphodiesterase 4 (PDE4) inhibitor (rolipram); or a large-conductance calcium-activated potassium channel blocker (charybdotoxin). The relaxation is expressed as percentage of preconstricted state, and data are presented as mean±s.e.m. Hyperoxic exposure significantly decreased the relaxation of TSM towards EFS, as compared to those obtained from control animals exposed to room air, and these impaired relaxant responses in hyperoxic preparations were significantly reversed in presence of VIP (p<0.01). VIP induced dose-dependent relaxation and the relaxant responses were overcompensated in hyperoxic animals. VPAC antagonist; PKA or PDE4 inhibition significantly reduced VIP-induced relaxation (p<0.05), while blockade of calcium-activated potassium channel did not have significant effect.The results of this study revealed that VIP reverses the impaired relaxation of airway smooth muscle induced by hyperoxic exposure via cAMP/PKA signaling pathway and we speculate that the use of VIP might be an effective therapeutic approach to prevent the airway hyperreactivity induced by hyperoxia. Supported by the Ministry of Education, Science, and Technology of the Republic of Kosovo - Small Research Grants: 2-2459-11. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.