bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ibuprofen causes mitochondrial and proteasome dysfunction in murine hearts
Authors: Sule Rasheed , Tiwari Shuchita , Rivera Gabriela , Gomes Aldrin 
Published: May 2023

Ibuprofen is one of the most commonly used compounds in the treatment of pain, fever, and inflammation. Despite its beneficial effects, the long-term use of ibuprofen has been previously associated with increased risks for cardiovascular diseases and all-cause mortality. Relatively little is known about the signaling pathways or mechanism(s) involved in ibuprofen’s ability to increase the risk of cardiovascular diseases. Very little is also known about sex-related differences with respect to ibuprofen. We investigated the effects of ibuprofen (100 mg/kg/day, equivalent to about a 500mg tablet/day for humans) on mouse hearts using proteomics. Proteomic results suggest that female mice hearts exposed to ibuprofen for 7 days had tenfold more changes than male mice hearts exposed to ibuprofen for 7 days. Both the expression and activity of β5i immunoproteasome was significantly increased in female mice heart relative to ibuprofen-treated male heart mice. Ibuprofen treatment altered mitochondrial function in both male and female mice. Ibuprofen also induced ROS generation and decreased mitochondrial membrane potential in both male and female mice. However, ibuprofen attenuated mitochondrial complex I activity only in female hearts. Several other signaling pathways were also altered only in female mouse hearts. This study found major ibuprofen-mediated selective effects on male and female mice hearts, with specific sex differences in mitochondrial and proteasome function in mice hearts. NIEHS/Superfund P42 ES004699 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.
Effect of bitter melon extract on cell survival and cell division markers in human metastatic breast cancer cells.
Authors: Bhetwal Bhupal , Remedi Marissa , Kopecek Kalie 
Published: May 2023

Introduction: Bitter melon extract (BME) is known to exhibit cytotoxic effects on breast cancer cells (MCF-7). However, the molecular mechanisms by which BME exerts cytotoxic effects are not established. Aims: We aimed to investigate if BME exerts cytotoxic effects on MCF-7 cells via inhibiting cell division and cell proliferation pathways. We hypothesized that BME inhibits proliferation of MCF-7 cells by decreasing activities of mitogen-activated protein kinase (MAP kinase) & cyclins. The MAP kinase and cylcins play critical roles in mitotic cell division and cell survival respectively. Methods: Fresh bitter melons were purchased from an Asian grocery store and the extract (BME) was extracted, centrifuged, and filter sterilized. The MCF-7 cells were cultured in DMEM medium with 1% BME (v/v). After culturing cells for 48 hours, pictures of cultures were taken to confirm cytotoxic effects on cells. In our earlier experiments, BME induced decrease in cell viability was also confirmed using Trypan blue exclusion and MTT assays. The cells were harvested and changes in the protein expression of Cyclin B1 and phosphorylated MAP kinase (P42/44, which is also known as Erk 1/2) were determined using anti-Cyclin B1 and anti- Phospho- Thr202/Tyr204 Erk1/2 antibodies in the western blotting. Results & Discussion: BME exerted cytotoxic effects on MCF-cells (N=4). BME almost completely wiped out the expression of cyclin B1 protein and phospho-Erk 1/2 (N=3 with duplicate gels). However, total Erk and GAPDH protein expressions were not affected by the BME treatment (N=3).The data together supports our hypothesis. In the future, we will be running more sets of experiments to confirm our findings. FRD-MUCOM. All authors contributed equally to this work. 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.
Multi-omic analysis of the cardiac cellulome defines a vascular contribution to cardiac diastolic dysfunction in obesity
Authors: Bender Shawn , Dona Malathi , Hsu Ian , Meuth Alex , Brown Scott , Bailey Chastidy , Aragonez Christian , Chandrasekar Bysani , Martinez-Lemus Luis , DeMarco Vincent , Grisanti Laurel , Jaffe Iris , Pinto Alex 
Published: May 2023

Coronary microvascular dysfunction (CMD) is associated with cardiac dysfunction and predictive of cardiac mortality in obesity, especially in females. Emerging evidence suggests development of heart failure with preserved ejection fraction in females with CMD and that mineralocorticoid receptor (MR) antagonism may be more efficacious in obese female, versus male, HFpEF patients. Accordingly, we examined the hypothesis that smooth muscle cell (SMC)-specific MR deletion prevents obesity-associated coronary and cardiac diastolic dysfunction in females. Obesity was induced in female mice via western diet (WD) feeding alongside littermates fed standard diet. Initial studies revealed that global MR blockade with spironolactone prevented impaired coronary vasodilation and diastolic dysfunction in obese females. Importantly, specific deletion of SMC-MR similarly prevented obesity-associated coronary and cardiac dysfunction. Cardiac gene expression profiling suggested reduced cardiac inflammation in WD-fed mice with SMC-MR deletion independent of blood pressure, aortic stiffening, and cardiac hypertrophy. Further mechanistic studies utilizing single-cell RNA sequencing of non-cardiomyocyte cell populations revealed novel impacts of SMC-MR deletion on the cardiac cellulome in obese mice. Specifically, WD feeding induced inflammatory gene signatures in multiple non-myocyte populations (B/T cells, macrophages, and endothelium), independent of cardiac fibrosis, that was prevented by SMC-MR deletion. Further, SMC-MR deletion induced a basal reduction in cardiac mast cells and prevented WD-induced cardiac pro-inflammatory chemokine expression and leukocyte recruitment. These data reveal a central role for SMC-MR signaling broadly impacting the cardiac cellulome and underlying obesity-associated coronary and cardiac dysfunction thus supporting the emerging paradigm of a vascular origin of cardiac dysfunction in obesity. Supported by NIH R01 HL136386 (Bender), NIH R01 HL119290 (Jaffe), the University of Missouri Research Core Facilities Grant Program (Bender), NHMRC Ideas Grant GNT1188503 (Pinto), VA Merit I01 BX004220 and RCS IK2 6004016 (Chandrasekar). The work was also supported by resources and the use of facilities at the Harry S. Truman Memorial Veterans Hospital in Columbia, MO. 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.
Antioxidant Flavonoid Diosmetin Is Cardio-Protective in a Rat model of Myocardial Infarction induced by Beta 1-Adrenergic Receptor Activation
Authors: Ahmad Taseer , Khan Taous , Kirabo Annet , Shah Abdul 
Published: May 2023

Myocardial infarction (MI), is a common and life-threatening manifestation of ischemic heart diseases. The most important risk factor for MI is hypertension. Flavonoids have been found to be efficacious in ischemic heart diseases by alleviating oxidative stress and beta-1 adrenergic activation but the mechanistic link is not clear. We hypothesized that antioxidant flavonoid diosmetin is cardio-protective in a rat model of MI induced by beta 1-adrenergic receptor activation. To test this hypothesis, we evaluated the cardioprotective potential of diosmetin on isoproterenol-induced myocardial infarction in rats by performing lead II electrocardiography (ECG), cardiac biomarkers including troponin I and creatinine kinase by using biolyzer 100, as well as histopathological analysis. We found that diosmetin (1 and 3 mg/kg) attenuated isoproterenol-induced elevation in T-wave and deep Q-wave on the ECG, as well as heart to body weight ratio (0.53 ± 0.01 vs. 0.34 ± 0.02, p< 0.001), and infarction size (55 ± 3.16 vs 13.33 ± 1.83 p< 0.001). In addition, pretreatment with diosmetin attenuated the isoproterenol-induced increase in serum troponin I (0.68 ± 0.012 vs 0.310 ± 0.0115, p< 0.001). These results demonstrate that flavonoid diosmetin may provide therapeutic benefit in myocardial infarction. 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.
Dipsogenic Role of Angiotensin II during Fear Memory Retrieval
Authors: Iyer Laxmi , Yu Zhe , Paronett Elizabeth , Swiercz Adam , Marvar Paul 
Published: May 2023

Motivational cues of thirst, sleep, and hunger impact memory and behavioral responses across species. Brain angiotensin II (AngII) is a potent thirst-producing peptide, where prior research demonstrated that angiotensin receptor blocker affects conditioned fear memory. We therefore hypothesized that acute water deprivation (WD) concurrent with angiotensin receptor blockage increases endogenous AngII levels and contributes to fear memory retrieval plasticity.C57BL/6J male mice underwent both auditory and contextual fear conditioning and received a single retrieval tone to initiate memory recall. During this time, mice were water restricted (24hrs) or ad-libitum in the presence/ absence of the angiotensin type1 receptor antagonist losartan (10mg/kg i.p.) given post fear memory retrieval. RT-qPCR, mass spectrometry, IHC, and ELISAs were performed to analyze specific synaptic plasticity and memory markers.Mice, which underwent WD during auditory memory retrieval, showed a significant reduction in freezing behavior in comparison to the non-WD controls during long-term memory (LTM) tests at 24hrs (p= 0.0019, n=12) and 1 week (p = 0.002, n=12). No difference in freezing response was observed in losartan-WD mice. Fear memory tests, after concurrent WD and contextual fear memory retrieval, showed no difference in freezing behavior between non-WD controls and WD with and without losartan (n=12). There was no significant difference in the baseline anxiety and locomotion behavior after water restriction. Following water deprivation, mass spectrometry revealed an elevated level of AngII peptide in the Subfornical region (SFO), an area essential for fluid regulation. The mRNA expression of RAS and synaptic plasticity genes - Agtr1, Agt and Bdnf (p<0.01, n=6) - were also significantly up-regulated only in WD saline mice. WD mice also had increased circulating aldosterone and angiotensin II (p<0.05, n=7) levels.Our findings indicate that elevated angiotensin II as a result of WD, reduced LTM, only during auditory fear memory reconsolidation, while altering the expression of specific RAS genes in SFO. Peripheral administration of losartan however blocked this effect. We propose that endogenous AngII as a dispogenic neuromodulator affects memory in a physiological state-dependent manner. 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.
Role of Rho kinase & Rhotekin in the bitter melon extract mediated cytotoxic effects on human metastatic breast cancer cells
Authors: Bhetwal Bhupal , Miller Claire , Kotlinski Henry , George Marie , Bleeke Jadyn 
Published: May 2023

Introduction: Bitter melon extract (BME) is known to exhibit cytotoxic effects on breast cancer cells (MCF-7). However, the molecular mechanisms by which BME exerts cytotoxic effects are not established. Aims: We aimed to investigate if BME exerts cytotoxic effects on MCF-7 cells via inhibiting Rho kinase (ROK) activity and Rhotekin expression. Both TOK and Rhotekin mediate several cellular processes such as cell division, attachment, migration etc. We hypothesized that BME exerts cytotoxic effects on MCF-7 cells by decreasing ROK activity and Rhotekin expression. Methods: Fresh bitter melons were purchased from an Asian grocery store and the extract (BME) was extracted, centrifuged, and filter sterilized. The MCF-7 cells were cultured in DMEM medium with 1% BME (v/v). After culturing cells for 48 hours, pictures of cultures were taken to confirm cytotoxic effects on cells. In our earlier experiments, BME induced decrease in cell viability was also confirmed using Trypan blue exclusion and MTT assays. The cells were harvested and changes in the protein expression of phospho-MYPT1 and Rhotekin were determined using anti-phospho-MYPT1 and anti- Rhotekin antibodies in the western blotting. Since ROK directly phosphorylates MYPT1, reduced expression of phospho-MYPT1 would indirectly suggest reduced ROK activity. Similarly, reduced expression of rhotekin is a downstream mediator of ROK activation pathway and reduced level of rhotekin would also contribute to the reduced viability of cells. Results & Discussion: BME exerted cytotoxic effects on MCF-cells (N=4). However, BME did not decrease expression levels of phospho-Thr 853-MYPT1 (N=3, duplicate gels). Also, BME did not affect Rhotekin expression (N=3, duplicate gels). Three different loading control proteins (actin, myosin lighta chain, and GAPDH) were used and none of them were affected by the BME. The data together rejects our hypothesis. In the future, we will be running more sets of experiments to confirm our findings. FRD-MUCOM. All authors contributed equally to this work. 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.
Pain and Plasticity: Spike Sorting Based Approach for Characterizing the Neural Circuitry Mediating Nociceptive Sensitization of the Defensive Strike Response in M. sexta
Authors: Valtierra Christian 
Published: May 2023

Nociception is the neural process of encoding and processing noxious stimuli; this sensation is transduced by specialized sensory neurons called nociceptors. Activation of nociceptors typically evoke so called “nocifensive behavioral responses,” that are generally aimed at decreasing exposure to the noxious stimulus. Intense activation of nociceptors can result in long lasting changes in the nervous system, as well as a subsequent decrease in the threshold for perceiving and acting upon stimuli as if it were harmful—this persistent change in the organism is nociceptive sensitization. The hornworm, Manduca sexta, displays rapid defensive “striking” behavior that accurately targets the cite of noxious stimulation of the abdominal body wall. Nociceptive sensitization of the defensive strike response has been induced behaviorally and recapitulated in vitro through extracellular electrophysiology, where it appears to be encoded by an increase in the firing frequency occurring within the central nervous system, rather than changes in peripheral nociceptor signaling. The current model proposed by Tabuena et al. (2017), argues that central neural alterations are responsible for the neural correlate of nociceptive sensitization (Tabuena et al., 2017), rather than changes in spiking activity of the primary sensory neurons. While the changes in firing frequency centrally are undisputed, relegating sensitization to the CNS alone may be overly simplistic, as the model is based on the comparison of threshold crossings of purported multiunit activity, rather than of single cell activity of known populations of sensory and/or higher order neurons. Moreover, studies in the vertebrate and invertebrate literature alike have reported retrograde sensitization of primary nociceptors (Parada et al., 2003; Xie et al., 2022; Babcock et al., 2000), mediated by inflammatory signals from the cite of injury, thus complicating the question of central versus peripheral sensitization. Therefore, it is worth re-examining the possibility of presynaptic plasticity as an added contribution to nociceptive sensitization. In this work, we propose leveraging available neuroanatomical data to re-analyze the Tabuena et al. dataset, by performing spike sorting and spike train analyses to tease apart the signals originating from the peripheral sensory neurons to obtain input/output relationships between the identified sensory units and VNC spiking during stimulation. Distinguishing the nociceptor and tracking its stimulus dependent firing is expected to elucidate the contribution of nociceptor activity in nociceptive sensitization. Genentech Scholars Foundation 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.
Skeletal muscle regeneration at different phases of the menstrual cycle
Authors: Ahmadi Mohadeseh , Pfeifer Brandon , Shimizu Krista , Hunter Werner , Altuhov Stepan , Nielsen Madeline , Hyldahl Robert 
Published: May 2023

INTRODUCTION: The effect of estrogen on muscle function and recovery from damage has been studied, but the effects of estrogen fluctuations during the menstrual cycle on muscle regeneration remain inconclusive. PURPOSE: The purpose of this study was to examine the effects of menstrual cycle phases on muscle regeneration after electrically stimulated muscle damage. We hypothesized that muscle damage during the late follicular (LF) phase results in better muscle regeneration as compared to muscle damage during the early follicular (EF) phase. METHODS: 23 premenopausal females (21.3±2 yrs) were recruited and split into two groups, EF and LF. After menstrual cycle tracking and phase confirmation, subjects underwent 200 electrically stimulated eccentric muscle contractions one week after providing a muscle biopsy. 7 days after inducing muscle damage, the subjects reported to give a final biopsy. Sections from muscle samples were stained for markers of laminin, dystrophin, satellite cells (SCs; Pax7), nuclei, total macrophages (CD68), and anti-inflammatory macrophages (CD206/CD68). Data for SCs, and %CD206+CD68+ cells are presented as mean±SD, and damaged fibers and CD206+CD68+ cells/fiber are presented as median±interquartile range. RESULTS: Electrical stimulation caused muscle fiber necrosis (0.005±0.018 damaged fiber/fiber, p=0.0002) as measured by the laminin-containing and dystrophin-deficient fibers. There was no difference between EF and LF groups for the number of damaged fibers (p=0.055) Muscle damage resulted in ~200% increase in the total number of CD68+ macrophages (p=0.008), and ~ 40% increase in the number of SCs (p=0.001) in both groups. However, there was no significant difference between EF and LF groups for the change in total macrophages per fiber (p=0.07) or SCs/fiber (p=0.25) in response to electrically stimulated muscle damage. Anti-inflammatory macrophages increased by about 104%±127 in EF and 240%±390 in LF group, but the difference between groups was not significant (p=0.39). A comparison of the proportion of anti-inflammatory macrophages per total macrophages before and one week after muscle damage did not show any time effect (p=0.25), which could indicate the regenerative and anti-inflammatory state of skeletal muscle one week after damage. CONCLUSION: Contrary to our hypothesis, our data indicate that skeletal muscle regeneration is not affected by different phases of the menstrual cycle. However, this finding is limited to only some of the markers of regeneration one week after muscle damage. 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.
Oxidative stress-related impairment of endothelial function in uncomplicated obesity
Authors: Kovarik Jack , Holwerda Seth 
Published: May 2023

Purpose: Visceral obesity is associated with vascular oxidative stress and development of hypertension. Therefore, we tested the hypothesis that suppression of vascular oxidative stress in adults with visceral obesity would improve endothelium-dependent dilation. Methods: IV infusion of a water-soluble antioxidant (ascorbic acid) was performed in 15 middle-age obese adults (3 men/12 women) without diabetes or hypertension in a placebo-controlled, double-blinded, crossover study design. Ascorbic acid was infused at 0.06g/kg fat free mass in 100 ml saline at 5 mL/min over 20 min (bolus) followed by a drip infusion at 0.02 g/kg fat-free mass in 30 ml saline at 0.5 mL/min over 60 min. Endothelium-dependent dilation was assessed via brachial artery flow-mediated dilation (FMD) and expressed as % increase in diameter from baseline before forearm occlusion. Results: Android fat distribution in the visceral cavity (VAT%) was 39 ± 10% and body mass index (BMI) was 37 ± 6 kg/m2. IV infusion significantly increased plasma ascorbic acid concentrations (Baseline: 42 ± 2 vs. Bolus: 66 ± 6 vs. End: 62 ± 4 μmol/L, P<0.001). Compared with baseline, bolus infusion of ascorbic acid significantly increased FMD (Baseline: 5.8 ± 2.5 vs. Bolus: 9.0 ± 4.0 vs. End: 8.0 ± 4.2 %, P=0.01), which remained after statistically adjusting for the stimulus (shear rate area under the curve) via analysis of covariance (P=0.04). No significant change in FMD was observed following placebo (saline) during a separate study visit (Baseline: 8.0 ± 4.4 vs. Bolus: 8.3 ± 3.9 vs. End: 8.6 ± 2.9 %, P=0.86). Conclusions: These preliminary findings suggest that acute elevations in plasma ascorbic acid concentrations improve endothelium-dependent dilation in middle-aged adults with visceral obesity. Future studies should target mechanisms by which mediators of oxidative stress in obesity (e.g., hyperlipidemia, hyperinsulinemia) inhibit nitric oxide-mediated vasodilation. Funding and support: R01HL159370-01 (S.W.H.). This work was also supported by a CTSA grant from NCATS awarded to the University of Kansas for Frontiers: University of Kansas Clinical and Translational Science Institute (# UL1TR002366). The contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or CTSA. 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.