bluuebunny/crossref_metadata_2025_split huggingface dataset:
Focally Acting Thrombin‐Inhibiting Nanoparticles Forestall Atherosclerosis in ApoE‐null Mice
Authors: Palekar Rohun U , Jallouk Andrew P , Myerson Jacob W , Pan Hua , Wickline Samuel A 
Published: November 2023

BackgroundThrombin plays a central role in mediating both thrombosis and inflammation through cleavage of its receptor, protease activated receptor 1 or PAR‐1. Activation of PAR‐1 results in the expression of numerous procoagulant and proinflammatory molecules involved with the development of atherosclerotic plaques. Herein, we describe the role of thrombin inhibition with anti‐thrombin nanoparticles in limiting plaque deposition and reducing inflammation and hypercoagulability in atherosclerosis.MethodsApoE−/− mice fed a Western Diet for 4 months, receiving three intravenous treatment doses per week for the penultimate 4 weeks of Western Diet feeding. Anti‐thrombin nanoparticles were used as the treatment consisting of perfluorocarbon nanoparticles conjugated to multiple copies of the direct thrombin inhibitor D‐phenylalanyl‐L‐prolyl‐L‐arginyl‐chloromethyl ketone (PPACK‐NP). Following 1 month of treatment, aortas were removed and the aortic arch was stained en face for deposited plaques using Sudan IV. Plaque burden in the aortic arch was quantified using ImageJ. Endothelial barrier disruption was quantified by measuring the deposition of semipermeant nanoparticles after 2 hours of circulation; and vessel procoagulant activity measured by time to carotid artery occlusion after dye‐laser injury.ResultsTreatment of ApoE−/− mice for one month with PPACK‐NP resulted in a 22.5% overall decrease (Fig. A) in aortic arch plaque burden compared to mice treated with saline (p = 0.03, Fig. B) in the face of no significant reduction in serum cholesterol (Fig. C). By immunofluorescence, both tissue factor expression and phosphorylation of NF‐kB p65 were reduced (25.94%, p = 0.027 and 21.62%, p = 0.017 respectively). Endothelial barrier function improved by 33% and procoagulant activity diminished by 46%. The activated partial thromboplastin time (APTT) normalized within ~30 minutes after bolus injection of nanoparticles confirming safety of the system with respect bleeding risk.ConclusionThese results demonstrate the ability of PPACK‐NP treatment to inhibit atherosclerosis in the plaque‐prone aortic arch, restore vascular barrier function, and reduce thrombotic risk without the need for cholesterol lowering or systemic anticoagulation.Support or Funding InformationResearch Support: NIH HL112303
Primary cilia of inner ear hair cells do not respond to mechanical stimuli
Authors: Indzhykulian Artur A. , Delling Markus , Liu Xiaowen , Li Yaqiao , Xie Tiao , Corey David P. , Clapham David E. 
Published: November 2023

Hair cells, the sensory cells of the inner ear, carry a bundle of 30–300 specialized actin‐based microvillus‐like projections called stereocilia. Tiny filamentous ‘tip links’ connect the stereocilia and transmit the force produced by sub‐nanometer movements to Ca2+‐permeable mechanoelectrical transduction (MET) channels at the tips of stereocilia—likely encoded by TMC1 and TMC2 genes.Adjacent to the tallest stereocilia is the kinocilium—a single nonmotile, (9+2) microtubule‐based true cilium. In chicken hair cells, kinocilia are connected to stereocilia by the same proteins as the tip links, raising the question of whether kinocilia also have MET channels. Indeed, TMC1 protein has been reported in kinocilia of mouse hair cells. In developing zebrafish hair cells, moreover, Ca2+ imaging experiments suggest that, prior to the onset of the normal MET, kinocilia mediate a mechanosensitivity elicited by reverse deflections of the hair bundle. Kinocilia and kinocilial links are required for a deflection‐dependent rise in cytoplasmic [Ca2+] in zebrafish.We used fast Ca2+ imaging to explore the mechanosensitivity of kinocilia on inner and outer hair cells of the mouse cochlea. We developed a transgenic mouse, Arl13b‐mCherry‐GECO1.2, which expresses a genetically‐encoded calcium indicator (GECI) in all primary cilia and, to a lesser extent, in stereocilia. We compared [Ca2+] changes in these two structures (stereocilia and kinocilia) of similar geometry (~5 μm length and 300–600 nm diameter). During hair‐cell development (E14‐P3), we used a swept‐field confocal with millisecond frame time to examine both hair‐cell kinocilia and primary cilia of nearby supporting cells (a negative control). With fast acquisition we could distinguish Ca2+ entry directly into cilia from Ca2+ that back‐diffused from cytoplasm. The fused mCherry and GECO fluorophores allowed ratiometric Ca2+ measurement. We observed robust deflection‐induced Ca2+ influx in stereocilia, but not in kinocilia, consistent with earlier measurements.Because kinocilia lack Ca2+ influx with motion, we went on to measure responses to flow in primary cilia in tissues upon which this hypothesis has been based: in cultured kidney epithelial cells, kidney thick ascending tubules, embryonic node crown cells, mouse embryonic fibroblasts, and MLOY‐4 and OCY454 osteocyte‐like cell lines. We found a complete lack of mechanically induced calcium increases in these primary cilia. Rapid Ca2+ influx specifically into cilia was not observed with physiological—or even superphysiological—levels of fluid flow in any of these cells, suggesting that primary cilia do not respond to mechanical forces via calcium signaling. We conclude that the Ca2+‐responsive mechanosensor hypothesis of primary cilia should be abandoned.Support or Funding InformationDPC: NIH 5R01 DC000304 and HHMI; DEC: HHMI.
Induction of Endothelium‐dependent Constriction of Large Arteries in Endotoxemic Hypotensive Rats
Authors: TSENG TZU‐LING , CHEN MEI‐FANG , LIU CHIN‐HUNG , PANG CHENG‐YOONG , HSU YUNG‐HSIANG , LEE Tony J.F. 
Published: November 2023

Effective managements of refractory hypotension to vasoconstrictors in severe sepsis are limited. A new strategy to ameliorate endotoxemic hypotension by inducing endothelium‐dependent constriction of endotoxemic large arteries was examined. Rat endotoxemia was induced by lipopolysaccharide (LPS, 10 mg/kg, iv). Changes of the hemodynamics were measured by in vivo experimentation, reactivity of isolated mesenteric arteries by blood‐vessel myography, and expression of proteins/enzymes by immunohistochemistry, biochemistry, and molecular biology. Six hours after LPS‐challenge, the diminished mean arterial pressure (MAP) was promptly reversed/normalized upon oroxylin‐A (OroA, a flavonoid from Scutellariae radix) injection (iv/ip). In isolated LPS‐treated but not normal mesenteric arteries, OroA (1–10 μM) induced endothelium‐dependent, sustained constriction. This vasoconstriction was blocked by endothelin‐1 (ET‐1)‐receptor antagonists, while OroA further enhanced LPS‐induced expression of endothelin‐converting enzyme (ECE), ET‐1 mRNA and proteins, and ET‐1 release, suggesting that OroA‐induced vasoconstriction was mediated by endothelial ET‐1. Also, OroA enhanced ROCK‐phosphorylation and reversed LPS‐induced suppression of RhoA‐activities in smooth muscle cells exclusively in arteries with endothelium. The activated muscle ROCK‐phosphorylation was blocked by ET‐1‐receptor antagonists and ROCK inhibitors. Moreover, OroA‐post‐treatment suppressed, via inhibiting NF‐κB activation, the inducible‐NOS expression and circulating NO. These results indicated that endothelial ET‐1 in endotoxemic mesenteric arteries, upregulated by LPS and released by OroA, induced prompt‐and‐sustained vasoconstriction via activation of muscle RhoA/ROCK‐pathway. In late endotoxemia, OroA‐induced vasoconstriction was caused partly by OroA‐induced decrease of circulating NO. The endothelium‐dependent constriction of large arteries induced by OroA supports its prompt reversal of diminished MAP. It is concluded that activation of endothelium‐dependent constriction of large resistance arteries and suppression of systemic inflammation offer new strategies for acute management of endotoxemic‐hypotensive shock.Support or Funding Information(Supported by MOST, Tzu Chi Foundation, & Tzu Chi University)
Cardioprotective effects of mitochondria‐targeted peptide SS‐20 in two different models of rat ischemia/reperfusion
Authors: Dai Wangde , Cheung Elissa , Alleman Rick J. , Brown David A. , Kloner Robert A. 
Published: November 2023

This study sought to determine effects of SS20 on myocardial infarct size in two different models of ischemia/reperfusion. For in vivo studies, Anesthetized Sprague Dawley rats were subjected to 30 minutes of coronary artery occlusion followed by 3 hours of reperfusion. Rats were randomized to control group (received saline infusion), low dose SS‐20 group (0.3 mg/kg/hour) and high dose SS‐20 group (3 mg/kg/hour) (n=15 rats in each group). Saline or SS‐20 were delivered into the jugular vein starting 5 minutes after coronary artery occlusion and were continued for one hour post coronary artery reperfusion. Body temperature, heart rate and blood pressure were monitored during the procedure. At the end of 3 hours reperfusion, ischemia risk area, no‐reflow area, and infarct size were measured. The ischemia risk area were similar among the saline (49.5 ± 2.3%), low dose SS‐20 (48.6 ± 2.1%), and high dose SS‐20 groups (48.7 ± 3.0%). Treatment with SS‐20 significantly reduced infarct size, which was expressed as % of risk area, in low dose (62.1 ± 4.4%) and high dose (64.0 ± 4.9%) compared with saline treatment (77.6 ± 2.6%, p=0.001 for both doses). There was no difference in infarct size between low and high dose SS‐20 treatment. The no‐reflow areas were comparable among the saline (23.9 ± 1.7%), low dose SS‐20 (23.7 ± 2.8%), and high dose groups (25.0 ± 2.1%). Body temperature, heart rate and blood pressure were comparable among the 3 groups at baseline, during ischemia, and at the end of 3 hours of reperfusion. In separate in vitro studies, isolated rat hearts were exposed to 20 minutes global ischemia, followed by SS‐20 administration (1μM) throughout the 2h reperfusion. Infarct size was reduced from 44 ± 3% in control group (n=11) to 17 ± 3% in the SS‐20 treatment group (n=5). There were no benefits of SS‐20 on recovery of left ventricular developed pressure, coronary flow, or maximal rates of contraction/relaxation. In summary, SS‐20 significantly reduced infarct size in two different models of myocardial injury, but did not affect hemodynamics or no‐reflow area in rat heart.Support or Funding InformationGrant support by Stealth BioTherapeutics Inc.
Occludin Degradation Mediated by Autophagy Leads to Blood‐Brain Barrier Disruption in Ischemic Stroke
Authors: Kim Kyeong‐A , Kim Eun‐Sun , Noh Dabi , Kim Eunji , Baek Seung‐Hoon , Bae Ok‐Nam 
Published: November 2023

The integrity of blood‐brain barrier (BBB), which shows selective permeability for substances to brain, is significantly damaged under ischemic stroke amplifying ischemic neuronal damage. There have been attempts to identify the exact mechanism of ischemic BBB disruption to minimize brain damage. Autophagy is a catabolic process which involves degradation and recycling of damaged or unnecessary organelles, but excessive autophagy can induce cell damage and death under pathological conditions such as ischemia. Here, we evaluated if autophagy plays a key role in BBB dysfunction under ischemic stroke. In vitro BBB model of bEnd.3 cells were exposed to oxygen‐glucose deprivation (OGD), an in vitro ischemic condition. After exposure to OGD for 18 hours, cell viability was significantly decreased and cellular permeability was impaired. The conversion of LC3‐I to LC3‐II and puncta of LC3 in bEnd.3 were increased, demonstrating that autophagy is induced under ischemic condition. Modulation of autophagy by 3‐methyladenine, an autophagy inhibitor, reversed the conversion of LC3, and restored the impairment of cell permeability induced by OGD. The protein level of occludin, a tight junction protein in BBB, was significantly decreased following OGD exposure, and reversed by 3‐MA. Also, immunofluorescence showed occludin was dispersed from cell membrane and co‐localized with LC3 puncta in bEnd.3 after OGD. Our findings showed that induction of autophagy might contribute to increased permeability through occludin degradation in brain endothelial cells under ischemia, providing a new mechanism of BBB disruption in ischemic stroke.Support or Funding InformationThis work was supported by grants from the Korean government (NRF‐2014R1A1A2058459 from the Ministry of Education and HI14C2284 from the Ministry of Health and Welfare).
Endothelial nitric oxide (eNOS)‐dependent and ‐ independent effects of hypoxia on vascular repair‐relevant functions of human CD34+ cells
Authors: Joshi Shrinidh , Jarajapu Yagna 
Published: November 2023

CD34+ hematopoietic stem cells are now known for their cardiovascular regenerative functions, and provide a breakthrough approach for the treatment of ischemic cardiovascular diseases. Vasorepairative functions of these cells are known to be potentiated by exposure to hypoxia. Previous studies have demonstrated that nitric oxide (NO) derived from eNOS plays an important role in the reparative functions of CD34+ cells. The current study evaluated the role of NO in the hypoxic potentiation of proliferation and migration, which are key functions that determine vasorepairative potential of these cells. Peripheral blood was obtained from healthy individuals (n=24) and mononuclear cells (MNCs) were separated by Ficoll centrifugation. MNCs were enriched for lineage negative cells, which were further enriched for CD34+ cells by immunomagnetic selection. Hypoxic exposure to hypoxia (1% oxygen) was accomplished by using a hypoxia chamber (Biospherix, Inc.). Intracellular NO levels were detected by DAF‐FM flow cytometry with and without the following treatments: Nω‐nitro‐l‐arginine‐methyl ester (L‐NAME), stromal derived factor‐1a (SDF) or vascular endothelial growth factor (VEGF). Effects of SDF or VEGF on proliferation and migration of CD34+ cells were evaluated under normoxic or hypoxic conditions. Hypoxic exposure increased intracellular NO levels (P<0.01, n=8) that were not affected by L‐NAME (300 μM) pretreatment (n=8). Basal proliferation and migration were enhanced (P<0.05, n=6) by hypoxic exposure that were resistant to L‐NAME (n=6). Proliferation and migration to either SDF or VEGF were potentiated by hypoxia (P<0.05, n=6), which was partly antagonized by L‐NAME (n=6). These findings suggest that hypoxia induces NO generation independent of eNOS, which potentiates proliferation and migration in CD34+ cells. Stimulatory effects of SDF or VEGF are at least in part mediated by eNOS activation in normoxic and hypoxic conditions.Support or Funding InformationThis work is partly supported by American Heart Association, and the Core Biology Facility at North Dakota State University, that was made possible by NIGMS (P30 GM103332‐01).
PARTICIPATION OF NO/GC/cGMP/K+ CHANNELS IN THE RELATATION INDUCED BY Crotalus durissus cascavella IN MESENTERIC ARTERY
Authors: de Jesus Rafael Leonne Cruz , Santos Suelen Silva , Simões Leticia Oliveira , Vasconcelos Walma , Veras Robson , Casais e Silva Luciana Lyra , Silvia Darizy Flavia 
Published: November 2023

IntroductionThe studies with toxins from snakes poison have shown promising biological effects, especially those regarding to cardiovascular system, such as hypotensive and vasodilators.AimsTo investigate the vascular effect of Crotalus durissus cascavella poison (CDC), a high prevalence species in the state of Bahia ‐ Brazil, was used rat superior mesenteric artery.MethodsWistar rats (12–16 weeks/250–300g) were euthanized in CO2 camera and the mesenteric artery were isolated and free from connective tissue and cut into rings that was kept in Tyrode solution at 37°C carbonated with 95% of O2 and 5% of CO2. The amperometry technic was used to evaluate NO production in isolated aortic endothelial cells. The significant values was p<0.05.ResultsCumulative administration of the CDC (0.001 – 30μg/mL) relaxed phenylephrine‐induced contractions in a concentration‐dependent manner (Emax = 47.9 ± 5.0%; n=8) in the endothelial vascular presence, this effect was practically abolished in the endothelial vascular absence (Emax= 5.8 ± 2.4%; n=5; *** p<0.001). The vasorelaxant effect induced by CDC in KCl 80mM‐induced contraction was abolished (Emax= 6.4 ± 0.9%; n=5; *** p<0.001. Pre‐treatment with Indomethacin (10μM), L‐NAME (100μM), and ODQ (10μM) significantly attenuated the relaxation effect induced by CDC (Emax = 29.1 ± 7.1%, n = 6, Emax = 17.5 ± 2.2%, n = 6, Emax = 11.2 ± 3.5%, n = 6, respectively). Additionally, in studies with isolated aortic endothelial cells demonstrated that CDC in concentration of 10 and 90 μg/mL induced significant increase in NO production. In mesenteric rings pre incubated with KCl 20mM, to evaluate K+ channels participation, the vasorelaxant effect induced by CDC was significantly reduced (Emax = 21.3 ± 8%, n = 7). Similar results was observed in the presence of Iberiotoxin (100nM), BaCl2 (30μM) and 4‐aminopiridin (1mM) [(Emax = 13.0 ± 3 %, n= 5, Emax = 21.7 ± 8 %, n= 5 e Emax = 9.5 ± 1.3, %, n=5, respectively).ConclusionThose results suggest that the CDC induces vasorelaxant effects in isolated superior mesenteric artery rings from normotensive rats in an endothelium‐dependent way, possibly trough endothelium‐derived relaxing factors, especially the NO, resulting in guanylyl cyclase activation, followed by GMPc production and probably K+ channels activation (BKca, Kv and Kir). Additionally, prostanoids derivated from cyclooxygenase also are involved in the relaxant effect induced by CDC.3012 ‐ ASPET Cardiovascular Pharmacology – General/OtherSupport or Funding InformationFundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Stimulation of the Immune System by Biophotonics Methods For Hepatitis B and Syphilis in an Inner City High Risk Population
Authors: Newton David , Einstein George P , Tulp Orien L 
Published: November 2023

Both Hepatitis B and Syphilis are sexually transmitted diseases of growing concerns in the U.S.A and globally. Syphilis affects over 12 million people in developing countries and between 700,000 and 1.6 million pregnancies a year. Hepatitis B is a main cause of acute and chronic liver disease in the United States and globally. Developing trends reveal that high‐risk heterosexual transmission is the leading cause of acute HBV infections in adults. Despite aggressive and innovative approaches to eliminate the prevalence of HBV and Syphilis, the medical community's best conventional approaches have not seen notable success rates. The objective of this study is to replace conventional medical approaches with Biophotonics to stimulate immune viability in patients affected with HBV and Syphilis among high‐risk urban population. The Literature Review will examine the immune viability of Biophotonics as a credible option to conventional medicine, explore the high prevalence of HBV and Syphilis in high risk urban setting; and examine the impact of these diseases and the failure of conventional approaches to fully reverse their prevalence. The participants in this study will include 30 men and women of varying ethnicities who have tested positive for HBV, Syphilis or both. Each participant will be treated with Biophotonics therapy and data will be interpreted via Quantitative Analysis. For effective therapy the G. P. Einstein Integrated Sigma Quanta Immunostimulation Energy Doses has been Applied. The findings will likely influence future therapeutic interventions to these diseases and trigger more research in the use of Biophotonics therapies.Support or Funding InformationSupported by Institutional Resources of USAT and the Einstein Medical Institute
Heterologous Phosphorylation of the Cannabinoid Type 1 receptor by Angiotensin II in Astrocytes isolated from Spontaneously Hypertensive Rats
Authors: Haspula Dhanush , Clark Michelle 
Published: November 2023

ObjectiveTo determine if a mediator of neuroprotection, the Cannabinoid Receptor Type 1 (CB1R), is phosphorylated by Angiotensin II (Ang II) via the Angiotension Type 1 receptor (AT1R) in astrocytes.BackgroundAT1R and CB1R have opposing roles in regulating the release of neuroinflammatory cytokines from astrocytes. While the former, a Gq G protein coupled receptor (GPCR), is pro‐inflammatory, the latter is anti‐inflammatory. Activation of Gq GPCRs have been demonstrated to inactivate the CB1R via a key phosphorylation event.MethodsAstrocytes were isolated from the brainstem and cerebellum of Spontaneously Hypertensive Rats (SHR), a model of hypertension and Attention Deficit Hyperactivity Disorder (ADHD). The cells were treated with 100 Nm Ang II for varying time periods and the extent of phosphorylation was measured using Western blotting and Flow cytometry. To determine the receptor involved, the cells were also pretreated with 10 μM Losartan and 10 μM PD123319, the AT1 and AT2 receptor antagonists respectively.ResultsBasal CB1R was significantly higher in SHR when compared to Wistar. Higher levels were observed in cerebellum. Ang II phosphorylated CB1R in both brain regions of both rat models. This effect was predominantly mediated through the AT1R. The phosphorylation event was initiated earlier and was sustained longer in SHR when compared to wistar.ConclusionHigher basal CB1R phosphorylation in SHR as opposed to Wistar is suggestive of a potential diminished protective effect in pathological conditions such as hypertension and ADHD. Also Ang II could be involved in disinhibiting CB1R actions via the AT1R by heterologous desensitization. Temporal and magnitude differences in phosphorylation of CB1R by Ang II, in SHR versus Wistar, are suggestive of a potential therapeutic target to combat neuroinflammation.Support or Funding InformationNSU HPD: Grant # 335585