bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reducing Thrombosis Risk With JNK‐2 siRNA Therapeutic Nanoparticles In ApoE−/− Mice
Authors: Pan Hua , Palekar Rohun U , Bacon John , Allen Stacy , Schlesinger Paul H , Wickline Samuel A 
Published: November 2023

IntroductionMacrophage foam cell formation is regulated in part by the MAPK family member, c‐Jun N‐terminal kinase, JNK2. Genetically modified ApoE−/− JNK2−/− mice are known to develop less plaque, suggesting that therapeutic strategies that inhibit JNK2 focally at the site of developing plaques could check the progression of atherosclerosis. However, whether JNK2 inhibition could modulate the risk of thrombosis in plaques is unknown. To this end, we report the utility of a new peptide‐based siRNA delivery nanoplatform that rapidly conveys serum‐stable anti‐JNK2 oligonucleotides into growing plaques (but not to or pass through normal vasculature) to inhibit plaque progression while simultaneously reducing plaque thrombotic risk.Methods and ResultsThe nanoconstruct designed to condense any siRNA into a 55nm particle comprised a nontoxic amphipathic cationic 21 a.a. peptide that facilitates both endosomal escape and release of the siRNA upon endosomal acidification. ApoE−/− mice at 6 weeks old were fed a Western diet for 14 weeks prior to treatment with either anti‐JNK2 siRNA NP (0.5 mg siRNA/kg; n=16) or HBSS (control; n=8) administered by intravenous injection twice weekly for 7 total doses under continued fat diet. Focal delivery of Cy5.5‐labeled siRNA was confirmed in excised aortas by fluorescence imaging 24 hours after nanoparticles injection. Real Time RT‐PCR and Western blots confirmed JNK2 knockdown in the total aorta in treated ApoE−/− at both mRNA (treated vs untreated aorta: 20% reduction p=0.034) and protein (treated vs untreated aorta: 50% reduction p=0.041) levels. Sudan IV staining of the arch revealed a 38% reduction in plaque surface area for control (47%) vs treatment (29%) groups (p=0.002). Thrombotic risk with or without JNK2 siRNA NP treatment was quantified by carotid artery dye‐laser injury according to occlusion times, indicating that JNK2 siRNA NP treatment increased occlusion time in treated mice by ~2× over that of control treatments, which confirms reduced procoagulant risk. Thrombin staining of the aorta sections demonstrated reduced thrombin deposition in plaque from the treated mice. Safety evaluations confirmed normal routine blood tests, and innate and adaptive immune responses.ConclusionAnti‐JNK2 siRNA nanoparticles specifically accumulate and are retained selectively in atherosclerotic plaque, inhibit plaque growth, and remarkably reduce thrombosis risk after only 3.5 weeks of biweekly therapy. In view of the exceptional safety profile, this therapeutic siRNA approach holds promise for rapidly stabilizing vulnerable atherosclerotic plaque.
Implementing Modified Active Learning Strategies in Graduate Pharmacology Education
Authors: Ponnoth Dovenia S. 
Published: November 2023

Active learning techniques and strategies are used to improve student learning while in class and can complement lecturing. Modifying select strategies and tailoring them to graduate education could be useful and creative pedagogical tools to encourage student learning and involvement in class. The aim of this study was to incorporate modified active learning approaches in several graduate pharmacology courses over a period of one year and evaluate in‐class student learning, participation and involvement. The different techniques employed included pre‐ and post‐lecture tests to solidify key concepts, team based learning to encourage discussion, increased use of the blackboard and modified concept maps to serve as learning tools. Each class (average class size was 15 graduate students) met once a week for 3 hours of contact time and along with the interactive exercises, traditional didactic lecturing was always part of the class. Other activities included journal clubs and discussion of peer‐reviewed articles. At the end of the year, an anonymous student survey was conducted not only to evaluate student learning and preferences but also use these responses to design future classes. The pre‐ and post‐tests were evaluated separately to assess learning. Each category on the survey could be scored a maximum of 4 points (strong agreement) and a minimum of 0 points (strong disagreement). The survey results showed students were not only enthusiastic about incorporating newer techniques into their learning but also had distinct preferences for particular exercises. Each category on the survey scored an average of 3.5 points, with the lowest score being 3.1 (use of the blackboard). The solving of modified concept maps was by far the most preferred activity (100% positive response, maximum average score 4) to aid learning. Based on the findings of this study, it was concluded that active learning can successfully be incorporated into graduate pharmacology education and using suitable exercises increases both student involvement and learning.Support or Funding InformationNA
Anti vascular inflammatory effect of Samul‐tang in HUVECs
Authors: Choi Eun Sik , Lee Yun Jung , Yoon Jung Joo , Ahn You Mee , Park Ji Hun , Han Byung Hyuk , Namgung Seung , Jin Xian Jun , Kang Dae Gill , Lee Ho Sub 
Published: November 2023

Samultang (, SMT) is a well known herbal prescription treating overall haematological disorders defined as blood deficiency, blood stasis and blood loss in traditional korean medicine. SMT is recorded in several formulary including ‘Treasured Mirror of Eastern Medicine’ (, Donguibogam) and consists of 4 herbs : Angelicae Radix, Cnidii Rhizmoa, Rhemanniae Radix Preparata and Paeonia Radix. Major cause of atherosclerosis and other vascular diseases is inflammation and vascular endothelium is emphasized as central spot of vascular inflammatory process. Thus, we investigated the effects of SMT water extracts on vascular inflammation in HUVECs (Human Umbilical Vein Endothelial Cells).Expression of CAMs (cell adhesion molecules) such as VCAM‐1 (vascular cell adhesion molecule‐1), ICAM‐1 (intracellular adhesion molecule‐1), E‐selectin (endothelial‐selectin) and HL‐60 monocyte adhesion were induced by TNF‐α stimulation. However, SMT pretreatment (10–50 μg/ml) inhibited CAMs expression and monocyte adhesion significantly. TNF‐α stimulation led HUVECs to produce ROS (reactive oxygen species) and translocate NF‐κB (nuclear factor‐κB) into nucleus, but SMT suppressed ROS production and NF‐κB nuclear localization significantly. Furthermore, activation of NF‐κB was also suppressed by SMT in dose dependent manner.HO‐1 (heme oxygenase‐1) protein level was significantly upregulated by SMT treatment (10–50 μg/ml) as HO‐1 inducer CoPP (cobalt protoporphyrin) did. Besides, nuclear translocation of Nrf‐2 (nuclear factor erythroid 2‐related factor 2), which transcripts antioxidative genes including HO‐1, was increased by SMT dose dependently. Intracellular NO (nitric oxide) and nitrite products were also increased by SMT treatment.Taken together, these results suggests that SMT might exerts vascular protective effects by suppressing vascular inflammatory process triggered from damaging stimulus including TNF‐α and by promoting synthesis of beneficial bioactive substances including HO‐1.
Effect of Hwangryunhaedoktang on TNF‐α‐induced vascular inflammation in human umbilical vein endothelial cells
Authors: Han Byung Hyuk , Lee Yun Jung , Yoon Jung Joo , Choi Eun Sik , Namgung Seung , Kim Hye Yoom , Ahn You Mee , Jin Xian Jun , Lee Ho Sub , Kang Dae Gill 
Published: November 2023

Hwangryunhaedoktang (HHT) originally recorded in an ancient Korea medicinal book named “Donguibogam” and has been used for the treatment of inflammatory hemorrhage relevance to vascular inflammation which can be cause an atherosclerosis. This study was designed to demonstrate whether HHT has an inhibitory effect on vascular inflammation induced by TNF‐α in human umbilical vein endothelial cells (HUVECs).Pretreatment with HHT decreased the adhesion of HL‐60 cells to TNF‐α‐induced HUVEC. HHT suppressed TNF‐α‐induced expression level of cell adhesion molecules such as intracellular adhesion molecule‐1 (ICAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1), and endothelial cell selectin (E‐selectin), as well as dose dependently inhibited TNF‐α‐induced matrix metalloproteinase‐2/‐9 activity. HHT significantly decreased TNF‐α‐induced intracellular reactive oxygen species (ROS) production. Western blot and immunofluorescence analysis showed that HHT inhibited the translocation of p65 NF‐κB to the nucleus. Phosphorylation of IκB‐α also was inhibited in cytoplasm by pre‐incubating with HHT, on the other hand increased expression of IκB‐α.These data showed that treatment with HHT regulate the ROS/NF‐κB signaling pathway. Therefore a traditional herbal formula HHT might be potential therapeutic agent of atherosclerosis.
H+ Induces Arteriolar Vasodilation but Does Not Initiate a Conducted Response in Skeletal Muscle Microvasculature
Authors: Charter Mackenzie E , Murrant Coral L 
Published: November 2023

Active hyperemia is the process where blood flow is recruited to a tissue to meet increased metabolic demand. Conducted responses are important to this process in order to direct blood flow to capillaries to feed active skeletal muscle fibres. A conducted response is a coordinated vasodilatory signal that travels from low to high branch order arterioles resulting in an increase in capillary perfusion. Conducted responses are hypothesized to be initiated by metabolic vasodilators ‐ byproducts of skeletal muscle metabolism‐ however, exactly which metabolites are involved remains unclear. Two vasodilatory metabolites of skeletal muscle metabolism are hydrogen ion (H+) and carbon dioxide (CO2). Both H+ and CO2 are interrelated through the bicarbonate (HCO3−) buffer (HCO3− + H+ <‐‐> H2CO3 <‐‐> CO2 + H2O) whereby H+ reacts with HCO3− to produce CO2 and CO2 reacts in reverse to produce H+. Thus, it is difficult to study the individual effects of these metabolites. In order to separate them we used a Tris (amine) buffer to superfuse the preparation, to allow us to apply H+ without changing CO2, thus allowing us to investigate the vasodilatory actions of H+ independently. Using an in situ, intact hamster cremaster muscle model, H+ was applied to the whole preparation through the superfusate or to a small region of a 2A arteriole (max. diameter approx. 40μm) via micropipette. Arteriolar diameter was recorded and analyzed offline. A range of H+ concentrations (63.1 – 251.2 nMol; pH 7.2 – 6.6) applied to the entire preparation induced arteriolar vasodilation. The maximum change in diameter was 9.9±2.1μm, which was not a maximal dilation in the arteriole (only 71.1% of maximal diameter). Micropipette application of 251.2 nMol H+ (pH 6.6), induced a local arteriolar vasodilation of 3.5±0.4μm however it did not elicit a significant vasodilation 1000μm upstream of the micropipette application site. These novel data demonstrate that H+ is a local arteriolar vasodilator with a limited capacity to cause vasodilation and does not cause a conducted response, which is important in directing blood flow to active skeletal muscle cells during active hyperaemia. These data bring into question the importance of H+ and changes in pH in the active hyperaemic response.Support or Funding InformationNSERC, Canada
Vascular Function Assessed by Passive Leg Movement and Flow‐Mediated Dilation: Evidence of Construct Validity
Authors: Rossman Matthew J , Groot H. Jonathan , Garten Ryan S , Witman Melissa , Richardson Russell 
Published: November 2023

The vasodilatory response to passive leg movement (PLM), often performed in either the upright seated or supine posture, appears to provide a novel, non‐invasive assessment of vascular function. However, PLM has yet to be compared to flow mediated vasodilation (FMD), a test recognized to have prognostic value in terms of vascular health. Therefore, to evaluate the construct validity of PLM, brachial (BA) and superficial femoral (SFA) artery FMDs (Doppler ultrasound) and upright seated and supine PLM were performed in 10 young (22±1) and 10 old (72±2) subjects. During PLM, leg blood flow (LBF, Doppler ultrasound), mean arterial pressure (MAP, Finapres), and leg vascular conductance (LVC, LBF/MAP) were measured, with peak change from baseline LBF (ΔLBF) and LVC (ΔLVC) characterizing the PLM‐induced hyperemic and vasodilatory responses. Age‐associated reductions in vascular function were evident with both PLM and FMD testing. During upright seated PLM, ΔLBF and ΔLVC were significantly correlated with BA (r=0.58 and r=0.69) and SFA (r=0.68 and r=0.64, for ΔLBF and ΔLVC, respectively) FMD. For supine PLM, there was some evidence of a positive relationship between both ΔLBF and ΔLVC and BA FMD (r=0.28 and r=0.3), but ΔLBF and ΔLVC were significantly related to SFA FMD (r=0.53 and r=0.55). Collectively, these data indicate that PLM, particularly upright seated PLM, likely provides a prognostic index of vascular health analogous to the traditional FMD test. Given the relative ease of PLM implementation compared to FMD testing, these data have important implications for the potential clinical use of PLM to assess vascular health.Support or Funding InformationNational Institutes of Health grant PO1 HL 09830, HL‐103786 and HL‐116579 and a VA Merit grant E6910R (RSR)
Sprague Dawley Severe Hemorrhagic Shock Model Assessed by Systemic and Microcirculatory Parameters
Authors: Nugent William H , Song Bjorn K 
Published: November 2023

Traumatic injury with hemorrhage is a primary cause of death in young, healthy adults with a significant portion of mortality occurring post‐resuscitation. Three factors contribute to outcomes: severity of hemorrhage, duration of hemorrhagic shock (HS), and resuscitation. To evaluate the efficacy of resuscitative intervention, we have developed a severe HS model sensitive to various techniques and solutions of resuscitation. Male Sprague Dawley rats were cannulated with arterial and venous lines to facilitate blood withdrawal, reperfusate infusion, continuous infusion of anesthesia, and monitoring of systemic variables (heart rate, mean arterial pressure [MAP]) with a pressure transducer (MP‐150; BIOPAC Systems Inc., Goleta, CA). The thin spinotrapezius muscle was exteriorized and secured in situ to a thermoregulated platform, which fit onto the stage of an upright microscope (Axio Imager.A2m, Carl Zeiss, Germany) for intravital microscopy of the microcirculation. Phosphorescence quenching microscopy assessed tissue oxygenation (ISF PO2). A 45% estimated blood volume withdrawal rate of 3.5 ml × kg−1 × min−1 produced a rapidly lethal (~1 h) model in sham (no treatment) animals with a good extension of survival times in animals resuscitated with either a non‐oxygen carrying colloid (Hextend™; Hospira, Inc, Lake Forest, IL) or whole blood. Hemorrhage produced a MAP of 20–25 mmHg and a non‐detectable ISF PO2. Following the 30 min HS phase, reperfusion of 20% estimated blood volume with Hextend™ elevated MAP to near baseline and increased survival times to 3 hours. However, Hextend™ did not change ISF PO2, which remained non‐detectable until the animals expired. Reperfusion of 20% estimated blood volume with whole blood elevated MAP back to baseline and elevated tissue ISF PO2 to normoxic levels. Additionally, whole blood extended survival times to 8 hours – the time of experiment termination. In summary, we have developed an in vivo model of severe hemorrhagic shock that allows for simultaneous characterization of systemic and microcirculatory metrics. The model is sufficiently severe to ensure consistency in post‐hemorrhage ischemia and mortality, while providing outcome sensitivity to reperfusion solutions and techniques.Support or Funding InformationSong Biotechnologies
Effects of Novel Oxygen Therapeutic on Systemic and Microcirculatory Variables in Sprague‐Dawley Rat Isovolemic Hemodilution Model
Authors: Song Bjorn K , Nugent William H , Sayes Christie M , Bruce Erica D 
Published: November 2023

Oxygen therapeutics (OTs), sometimes referred to as blood substitutes, have been investigated for almost half a century, and remain actively researched. OT's have traditionally been categorized as either hemoglobin‐based oxygen carriers (HBOCs) or perfluorocarbons (PFCs). Unlike blood, HBOCs and PFCs do not require blood typing, have a long shelf life, do not transmit blood borne diseases, and in most cases do not need refrigeration. Despite the promising attributes of HBOCs and PFCs, there are concerns regarding arteriolar constriction that leads to increased blood pressure and increased white blood cell content for HBOCs and PFCs, respectively. This study investigated the systemic (heart rate, mean arterial pressure) and microcirculatory (vasoactivity, interstitial fluid oxygenation ‐ISF PO2) impact of a novel OT named OX66 (Hemotek LLC, Plano, TX). OX66 is different from HBOCs and PFCs in that it does not contain hemoglobin or long carbon chains; rather, it is a poly‐oxygenated aluminum hydroxide that contains 66.20% oxygen. OX66 is a true clathrate (lattice‐like) structure having large areas within the structure able to capture and hold oxygen. In this study, male Sprague‐Dawley rats underwent a 50% blood volume isovolemic hemodilution exchange with either OX66 or phosphate buffered saline (PBS; volume control), since OX66 was suspended in PBS. Isovolemic hemodilution is the reduction of red blood cells (hematocrit) with an equal volume of hemodiluent, i.e., crystalloids, colloids or OTs. The withdrawal/infusion rate was 2.0 ml × min−1 × kg−1 and performed through a cannulated carotid artery and jugular vein. Systemic measurements were recorded via a cannulated femoral artery that was connected to a pressure transducer (MP150; Biopac Systems, Inc, Goleta, CA), while microcirculatory parameters were collected through phosphorescence quenching and intravital microscopic examination of the exteriorized spinotrapezius muscle. Compared to baseline, a 50% blood volume exchange with either hemodiluent produced a significant reduction in heart rate, blood pressure, arterial diameter and ISF PO2 in all animals. However, OX66 animals demonstrated an improvement in ISF PO2 compared to PBS animals. This finding demonstrates that OX66 both transports and releases oxygen to the peripheral microcirculation.Support or Funding InformationSupport: Baylor University‐Hemotek Medical Applications Project: 32370181
A Dynamical Computational Model of Circulating Blood Gases, Hemoglobin Binding, and pH Regulation in the Microcirculation
Authors: Bassingthwaighte James B , Dash Ranjan K. 
Published: November 2023

A new computational model is developed to characterize the dynamical aspects of simultaneous blood‐tissue transport and exchange of O2, CO2, HCO3− and H+ in the coronary microcirculation by accurately simulating the rapidly changing intra‐capillary concentration profiles as the blood gets warmed, acidified, deoxygenated, and carbonated while flowing through the myocardial capillaries. It is based on our 2006 steady‐state, nonlinear, convection‐diffusion‐permeation‐reaction computational model of circulating blood gases, hemoglobin (Hb) binding, and acid‐base buffering in the blood which is available for download from our JSim model repository (http://www.physiome.org/jsim/models/webmodel/NSR/Exchange_O2_CO2_HCO3_and_H+). The kinetics are complicated by: (i) intravascular axial diffusion, (ii) RBC/plasma velocity ratio > 1, (iii) gas and ion permeation through RBC, endothelial and cell membranes, (iv) carbonic anhydrase‐catalyzed CO2 hydration reaction and HCO3‐ buffering in blood and tissue, (v) nonlinear competitive temperature‐dependent binding of Hb, O2, CO2 and H+ inside RBCs, (vi) myoglobin‐facilitated transport of O2 in cardiac cells, and (vii) O2 consumption and CO2 production inside mitochondria, with CO2 production and the respiratory quotient dependent upon the substrates, gas concentrations, cardiac work levels, and the rate of mitochondrial oxidative phosphorylation. These interactions smooth the axial spatial gradations in fluxes of O2 from blood into tissue and of CO2 out into the lung air. The simplest version of this extended model focuses on intracapillary RBC‐plasma interactions and the conditions for alveolar‐arterial differences in oxygen saturation. The cell‐tissue versions incorporate a new kinetic model of cytochrome c oxidase (CcO) that accurately characterizes the kinetics of O2 consumption by CcO as a function of cytochrome c redox fraction, mitochondrial inner membrane electrical and proton gradients (i.e. proton motive force; energy state), O2, and pH. The model serves as the dynamical exchange component of whole body gas exchange and acid‐base regulation. With augmentation to bind 15O2 or 17O2, this model can be used for the analysis of dynamic PET image sequences or NMR studies for estimating the regional tissue O2 consumption.Support or Funding InformationSupported by NIH grant U01‐HL122199
Rho‐Kinase Inhibition Restores ATP Release from Deoxygenated Red Blood Cells of Older Adult Humans
Authors: Racine Matthew L. , Joyner Michael J. , Dinenno Frank A. 
Published: November 2023

Substantial evidence indicates that red blood cells (RBCs) can act as a ‘sensor’ for oxygen demand in addition to their traditional role as an oxygen ‘carrier’, in that they are able to stimulate vasodilation and increase oxygen delivery to the tissue through the release of ATP in direct proportion to the degree of hemoglobin (Hb) deoxygenation. However, RBCs from older adults have an impaired ability to release ATP in response to Hb deoxygenation compared to RBCs from young adults, yet the underlying cause of this remains unknown. RBC membrane deformability has been shown to decrease with advancing donor age, thus we hypothesized that increasing membrane deformability (via the Rho‐kinase inhibitor Y‐27632) would restore ATP release from RBCs of older adults, while decreasing membrane deformability (via the cell‐stiffening agent diamide) would impair ATP release from RBCs of young adults. Isolated RBC ATP release in response to normoxic (PO2 ~110 mmHg) and hypoxic (PO2 ~20 mmHg) stimuli was quantified in RBCs from young (23 ± 3 years; n = 11 for Y‐27632 and diamide) and older (68 ± 7 years; n = 11 and 6 for Y‐27632 and diamide, respectively) adults using the luciferin‐luciferase technique following RBC incubation with either saline (control), Y‐27632, or diamide. The relative change in ATP release from normoxia to hypoxia in saline control conditions was impaired in RBCs from older (41.5 ± 12.1% and 63.5 ± 18.5%) vs. young (110.4 ±12.7% and 146.0 ± 29.3%) adults (P < 0.05; control trials for Y‐27632 and diamide, respectively). Y‐27632 improved RBC ATP release in older adults (98.8 ± 19.8%) such that it was not different from the young control (P > 0.05), whereas diamide decreased RBC ATP release in young adults (57.4 ± 8.4%) such that it was not different from the older control (P > 0.05). Preliminary data from our laboratory indicate that these drugs affected RBC membrane deformability as expected based on other reports of the effects of Y‐27632 and diamide on RBC membrane deformability. We conclude that decreased membrane deformability contributes to the impaired ability of RBCs from older adults to release ATP in response to Hb deoxygenation, and that this may have implications for impaired vascular control with advancing age.Support or Funding InformationHL095573, HL119337, F31HL126377