bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Platelet Aggregation, Secretion, Integrin Activation, and Calcium Release Stimulated by Different Agonists in Native Americans
Authors: Williquett Brady , Valen Grace E. , Goerger Krista , Clarke Santa , Abbott Karla , Larson Mark K. 
Published: April 2020

Arterial thrombosis results from obstructive vascular blood clots that are largely initiated by platelet activation. As such, individuals whose platelets react more robustly to chemical signals generated by vascular damage or dysfunction are at a higher risk for morbidity and mortality caused by thrombosis. As a population, Native Americans have higher rates of obstructive arterial diseases, and previous research suggests African American populations have higher overall platelet reactivity that positively correlates to their higher risk for cardiovascular disease. Therefore, we set out to measure platelet reactivity in Native Americans and identify potential genetic alleles that correlate to the elevations in responsivity. Five platelet agonists were utilized to simulate vascular damage signals, followed by measurements of markers indicative of subsequent platelet activation: aggregation, secretion, integrin activation, and calcium ion mobilization. Preliminary results from 17 subjects showed that Native American platelets more rapidly aggregate in response to several agonists when compared with Caucasians, whereas other measures of platelet function are largely similar between the groups. These results suggest that Native Americans have more sensitive platelet aggregation responses, which could result in a higher risk for occlusive arterial diseases. Complete genomic sequences were also obtained from the highest responders for use in identifying alleles that correlate to this response.Support or Funding InformationInstitutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103443
Cancerous Phenotypes Exhibited by NIH/3T3 Fibroblasts Overexpressing BST‐2
Authors: Alvarez Folnetti , Okeoma Chioma 
Published: April 2020

Oncogenesis is a multistep process initiated by the accumulation of genetic and/or epigenetic changes that culminate in the transformation of normal cells into cancer cells. The genetic and/or epigenetic changes result in dysregulated gene expression signature that may involve key cancer driving genes such as Bone Marrow Stromal Antigen 2 (BST‐2). In this study, we sought to test the hypothesis that BST‐2, a type II transmembrane protein regulates early events of oncogenesis. BST‐2 is a multifaceted protein with both antiviral and proviral functions and has been implicated in promoting aggressive cancer cell behavior such as migration and invasion. Meta‐analysis of breast cancer datasets from The Cancer Genome Atlas (TCGA) show a statistically significant correlation between elevated BST‐2 expression and poor survival. In addition to its role in breast cancer progression, elevated BST‐2 expression correlates with poor prognosis in endometrial, brain, lung, and oral cavity cancers, further implicating elevated BST‐2 expression in cancer progression. However, the role of BST‐2 in regulating early oncogenic events is not well understood. Our approach consisted of stably overexpressing exogenous human BST‐2 in NIH/3T3 mouse embryonic fibroblasts that express very low endogenous mouse BST‐2 to generate 3T3BST‐2 cells. The control cells were stably transfected with pCDNA3.1 to generate 3T3pCDNA cells. The cells were tested for phenotypes associated with early oncogenic events under two‐dimensional (2D) and three‐dimensional (3D) culture conditions. In a confluent 2D monolayer, 3T3BST‐2 cells compared to 3T3pCDNA cells resisted contact inhibition and had enhanced growth and survival that is associated with increased phosphorylated protein kinase AKT (T308), increased phosphorylated mitogen‐activated protein kinase (ERK1/2), and decreased pro‐apoptotic factor Bak. In a 3D spheroids model, 3T3BST‐2 spheroids were larger in area and had higher ATP content, an indication of anchorage‐independent survival. Suspended as single cells in a gel, 3T3BST‐2 cells also had enhanced survival, especially when arginylglycylaspartic acid (RGD) adhesive peptide motif was present. Suppressed contact inhibition and enhanced anchorage‐independent survival suggests that overexpression of BST‐2 contributes to malignant transformation of normal cells. Discerning the mechanisms of BST‐2 mediated oncogenesis will expand our understanding of events that transforms a normal cell into an unwanted cancerous cell, which may potentially form the foundation for the development of BST‐2‐targeted cancer therapies.Support or Funding Information1. Molecular and Cellular Pharmacology PhD program 2. CIE’s IMSD‐MERGE scholars program 3. Scholars in Biomedical Sciences program (SBMS) 4. Stony Brook University Cancer Center
Canagliflozin Inhibits Vascular Smooth Muscle Cell Proliferation and Migration
Authors: Durante Giovanna L. , Khanna Yash P. , Behnammanesh Ghazaleh , Peyton Kelly J. , Durante William 
Published: April 2020

Cardiovascular disease is the major cause of morbidity and mortality in diabetes. Although numerous factors contribute to the development of diabetes‐associated cardiovascular disease, excessive proliferation and migration of vascular smooth muscle cells (SMCs) plays a critical role. Sodium‐glucose cotransporter 2 (SGLT2) inhibitors are the latest approved class of glucose lowering drugs. By blocking glucose uptake in the proximal tubule of the kidney, SGLT2 inhibitors induce glycosuria leading to decreases in fasting and postprandial glycemia. Notably, large multicenter clinical trials have demonstrated that SGLT2 inhibitors reduce cardiovascular disease and mortality in patients with type 2 diabetes. The mechanism underlying the cardiovascular benefits of SGLT2 inhibitors is not fully known; however, direct effects on vascular SMCs have not been considered. In the present study, we investigated the effect of three different SGLT2 inhibitors on vascular SMC function. Treatment of rat and human aortic SMCs with canagliflozin resulted in a potent, concentration‐dependent inhibition of cell proliferation and DNA synthesis. The anti‐proliferative effect of canagliflozin was observed at clinically relevant concentrations, and this was not seen with other SGLT2 inhibitors, including empagliflozin and dapagliflozin. The inhibition of SMC growth by canagliflozin occurred in the absence of cell death and was associated with the arrest of SMCs in the G0/G1 phase of the cell cycle. In addition, scratch‐wound assays found that vascular SMC migration was blocked by canagliflozin in a concentration‐dependent manner, whereas empagliflozin or dapagliflozin had no effect on SMC motility. In conclusion, the present study identified canagliflozin as a robust inhibitor of vascular SMC proliferation and migration. These inhibitory effects of canagliflozin are detected at pharmacologically relevant concentrations and are unique for this drug and not seen with other SGLT2 inhibitors. The ability of canagliflozin to exert these pleiotropic effects on vascular SMC function may contribute to the salutary cardiovascular actions of this drug in patients with type 2 diabetes.Support or Funding InformationSupported by American Diabetes Association grant #1‐17‐IBS‐290.
A MAP kinase‐driven mechanical stress response mediates the hypertrophic response in skeletal muscle
Authors: Nordgaard Cathrine , Blasius Melanie , Vind Anna Constance , Reinert Marie Sofie , Snieckute Goda , Tiedje Christopher , Bekker-Jensen Simon 
Published: April 2020

When cells are exposed to unfavorable conditions they cope by activating cellular stress responses. Such signaling depends on activation of the p38 and JNK MAP kinases and is crucial for both general tissue homeostasis and a range of physiological reactions. However, the mechanisms linking diverse stress stimuli to MAP kinase activation remain poorly understood.Here we show how a specific MAP3 kinase mediates activation of p38 and JNK upon hyperosmotic‐ and mechanical stress. Employing fluorescence microscopy techniques, we show that this MAP3 kinase translocates from the cytoplasm to the nuclear lamina and other detergent‐resistant structures in the nucleus upon osmotic stress induction. Here, it co‐localizes with structures rich in DNA damage response factors such as 53BP1, but void of DNA strand breaks, which were shown to constitute nuclear sensor domains for mechanical stress.Mechanical stress is experienced in skeletal muscle due to the repeated contraction of muscle fibers upon physical exercise. The signaling from these events underlie the hypertrophic response, where muscles respond to exercise by growing larger and stronger. We show that the above signaling pathway operates during exercise‐associated stimulus of C2C12 myotubes, where it is responsible for the production of a range of inflammatory transcripts and myokines (such as IL6). Consequently, in a mouse model deficient for the MAP3 kinase, we see clear signs of pathology associated with decreased muscle repair and adaptation to exercise.In conclusion, our work provides novel insight into the mechanisms by which cells sense and react to mechanical stress stimuli and identify skeletal muscle as a major target organ upon deficiency in these responses.
TRPC3 is a critical contributor to the osmosensitivity and renal water handling in the mouse collecting duct
Authors: Tomilin Viktor N. , Mamenko Mykola , Zaika Oleg , Ren Guohui , Pochynyuk Oleh 
Published: April 2020

Kidney plays a central role in control of systemic water balance depending on hydration status. Arginine vasopressin (AVP) augments water permeability of the collecting duct (CD) by facilitating aquaporin 2 (AQP2) translocation to the apical plasma membrane. Inability of CD to respond to AVP signal causes Nephrogenic Diabetes Insipidus (NDI) leading to polyuria, dehydration and thirst. AVP stimulates CD to reabsorb water, but this can only occur in presence of positive osmotic difference between the cytosol and luminal fluid. Extracellular hypotonicity increases [Ca2+]i and causes cell swelling due to AQP2‐driven water influx. The significance of the osmosensitive [Ca2+]i signaling for renal water transport and urinary concentration remains unknown. TRPC3 is a Ca2+ permeable mechano‐activated channel abundantly expressed in the CD. Using ratiometric Fura 2‐based [Ca2+]i imaging, we simultaneously measured [Ca2+]i dynamics and the rate of cell swelling as a readout of the AQP2‐dependent water reabsorption in freshly isolated split‐opened CDs of wild type and TRPC3 −/− mice and combined it with immunofluorescent detection of AVP‐induced AQP2 trafficking and assessment of systemic water balance. We found that TRPC3 inhibition or deletion precluded [Ca2+]i elevations induced hypotonicity and remarkably slowed the rate of cell swelling indicative of diminished water transport in the CD. TRPC3 −/− mice had comparable with WT serum and urine osmolarity in the presence of ad libitum water intake, but exhibited a significantly greater bodyweight loss, and larger urinary volume after 24 h water deprivation despite higher AVP levels when compared to WT. Furthermore, osmosensitive [Ca2+]i elevations were greatly increased in CDs from WT but not in TRPC3 −/− animals after 24 h water deprivation. The rate of cell swelling was greatly accelerated in WT, while was only modestly increased in TRPC3 −/− mice under this condition. Using immunofluorescent microscopy, we found that AQP2 translocated to the apical plasma membrane in WT, while remained largely cytosolic in TRPC3 −/− after 24 h water deprivation. Consistently, TRPC3 inhibition interferes with AVP‐induced AQP2 trafficking in cultured mpkCCDc14 cells. In summary, we demonstrate a significant role of TRPC3 in osmosensitivity and regulation of AVP‐dependent AQP2 trafficking in the CD. TRPC3 deletion compromises systemic water balance producing an NDI‐like phenotype.Support or Funding InformationAmerican Heart Association Career Development Award for Viktor Tomilin (19CDA34660148)NIH RO1 awards for Oleh Pochynyuk (DK117865 and DK119170)
Integration of Gross Anatomy Laboratory sessions into Medical Physics Curriculum using Segmentation and Augmented‐Reality
Authors: Noel Geoffroy P.J.C. , Kang Esther S.H. , Popovic Marija 
Published: April 2020

Anatomy teaching to Medical Physics (MP) trainees is recognized by the Commission on Accreditation of Medical Physics Education Programs, and yet anatomy laboratory sessions have not been integrated in MP training curricula. This study outlines the development of anatomy laboratory sessions for MP trainees and serves as the initial step to developing an anatomical curriculum guideline for MP. The two objectives of this qualitative study are: (a) to explore the educational potential of integrated anatomy laboratory sessions in the MP curriculum and (b) to evaluate the benefits of interprofessional education activities between MP trainees and Radiation Oncology residents.Participants included 17 MP graduate students, 2 MP residents and 8 Radiation Oncology residents, over two years. Two 2‐hour anatomy laboratory sessions per year were organized following the respective in‐class lectures on the pelvis, head and neck and thorax. After the two laboratory sessions, participants were asked to fill out a survey reflecting on their experience in the laboratory.Both MP students and radiation oncology residents voiced an appreciation for the anatomy laboratory sessions for the consolidation and application of knowledge. Most mentioned that these sessions helped to “visualize” and “have a better understanding” of the anatomy taught in class and made them more “confident”. In terms of interprofessionalism, both the MP and the radiation oncology trainees expressed a better understanding of one another’s level of knowledge in anatomy. Both groups suggested to have more interprofessional sessions. They also suggested to build a stronger link between cadaveric anatomy and imaging anatomy through augmented reality technology.The integration of anatomy laboratory sessions into the anatomy curriculum for MP enhances trainees’ understanding of human anatomy and its pivotal role in radiation therapy treatment. Furthermore, interprofessional activities prior to clinical placement of MP students reinforce technical and professional communication.Support or Funding InformationThe authors would like to thank the support provided by the Centre for Medical Education Innovation and Research Seed Fund (to GPJCN).
Dendritic Cell Specific Jak2 Expression plays a role in in Salt‐Sensitive Hypertension through an ENaC‐dependent Mechanism
Authors: Aden Luul A. , Pitzer Ashley A. , Grimes Samantha , Ray Evan , Kleyman Thomas , Kirabo Annet 
Published: April 2020

Salt‐sensitivity of blood pressure affects 50% of hypertensive and 25% of normotensive individuals and is an independent predictor of death due to cardiovascular disease. We recently found that gamma and alpha subunits of the epithelial sodium channel (ENaCαγ) on dendritic cells mediate NADPH oxidase‐dependent formation of immunogenic isolevuglandin (IsoLG)‐protein adducts leading to inflammation and salt‐sensitive hypertension. We hypothesized that Jak2 expression in dendritic cells regulates expression of ENaCγ and promotes salt‐sensitive hypertension. Mice lacking Jak2 in CD11c+ cells developed blunted hypertension (123.8±4.7) during the high salt feeding phase of the N‐Nitro‐L‐arginine methyl ester hydrochloride (LNAME)/high salt model of salt‐sensitive hypertension compared to their wildtype littermate controls (140.5±6.5). These mice also exhibited less infiltration of monocyte/macrophages in their kidneys and less volume retention (69.55±5.8) in response to high salt‐feeding when compared to the wildtype litter mate controls (57.89±9.5). We also found that deletion of Jak2 in dendritic cells reduced the salt‐induced expression of ENaCγ in CD11c+ cells. Following high salt feeding, mice lacking Jak2 in DCs exhibited less aortic infiltration of CD11c+ cells with less expression of CD86 and production of IsoLGs. These results indicate that dendritic cell Jak2 plays a role in salt‐sensitive hypertension through an ENaC‐dependent mechanism.Support or Funding InformationK01HL130497R01HL147818T32HL144446
Pharmacokinetic studies of moracin C in mice using various blood sampling methods
Authors: Choi Young-Hee , You Byung-Hoon , Chin Young-Won 
Published: April 2020

Mice have been increasingly used in pharmacokinetic evaluation due to using less amount of compound and availability of various disease models and blood sampling methods in mice have produced the distinct pharmacokinetic profiles even in the same compounds. In this study, three blood sampling methods [i.e., automated dosing/blood sampling (ABS), tail artery collection (TA) and heart puncture (HP)] were applied to investigate the pharmacokinetics of moracin C in mice. Depending on blood sampling sites, significant differences were observed in plasma concentrations as well as pharmacokinetic parameters such as AUC (area under the plasma‐time concentration curve). The AUC values in ABS and TA groups were smaller than that in HP group. Considering that moracin C was greatly distributed to liver and gastrointestinal tracts (e.g., stomach, small intestine, and large intestine) and was extensively metabolized via glucuronidation in liver and small intestines, the AUC differences could be mainly caused by the stronger stress from blood sampling via heart puncture, which might interfere the elimination (i.e., hepatic and/or intestinal first‐pass effects). The remaining amounts of moracin C in gastrointestinal tract at 24 h after its oral administration were similar among three groups with different blood sampling methods, indicating that the absorption of moracin C was not affected during the blood sampling stress of HP. These results indicate that the ABS or TA method can be utilized to obtain better reliable and accurate pharmacokinetic parameters in mice.Support or Funding InformationThis study was supported by National Research Foundation of Korea (NRF) grants funded by the Korea government (MSIT) (NRF‐2016R1C1B2010849, NRF‐2019R1A2C2009053 and NRF‐2018R15A2023217).
A nigro‐vagal pathway controls colonic motility and may be impaired in a model of environmental Parkinson’s disease
Authors: Nanni Giorgia , Travagli R. Alberto 
Published: April 2020

Parkinson’s disease (PD) is a neurodegenerative disorder characterized by motor deficits caused by the loss of substantia nigra pars compacta (SNpc) dopaminergic neurons. It is well known that the majority of PD patients experience gastrointestinal (GI) dysfunctions such as gastroparesis, impaired swallowing and severe constipation. Upper GI tract functions are modulated by vagal reflexes, the central neurocircuit of which are located in the dorsal vagal complex (DVC, i.e. dorsal motor nucleus of the vagus (DMV), nucleus tractus solitarius and area postrema). We discovered a monosynaptic nigro‐vagal pathway recently that modulates gastric tone and motility and is impaired in an environmental PD model induced by oral administration of subthreshold doses of paraquat (P) and lectin (L). It is not known, however, whether this pathway also modulates colonic motility and whether the P+L model of PD decreases intestinal transit.Proximal colonic motility and tone were recorded with a strain gauge aligned with the colonic circular smooth muscle, approximately 3 cm distal to the ileal‐cecal valve. Microinjections of the ionotropic glutamate selective agonist, NMDA, were made at the level of the SNpc (5nmoles/210nL) and the effects on colonic motility and tone assessed. Microinjection of NMDA, but not vehicle, increased colonic tone (472±54mg) and motility (+358±166% of baseline; N=2).Gastro‐cecal transit, an indicator of intestinal motility, was measured using the 13C lactose ureide‐ breath test. Female rats had a slower gastro‐cecal transit time (178±24 min, N=4) than male rats (138±1.5 minutes, N=3). Treatment with P+L delayed gastro‐cecal transit time to 272±18 minutes (N=3).These data suggest that the nigro‐vagal pathway also modulates colonic motility and the P+L model of PD delayes intestinal transit and thus may be a useful model to study PD‐related constipation.Support or Funding Informationsupported by NIH grant DK‐55530 to R. Alberto Travagli
Nox4‐mTORC1 connection in salt‐induced hypertension and kidney injury in Dahl S rats
Authors: Kumar Vikash , Geurts Aron , Kurth Theresa , Zheleznova Nadezhda N. , Yang Chun , Cowley Allen W. 
Published: April 2020

Chronic treatment of Dahl salt‐sensitive (SS) rats with rapamycin which specifically inhibits mTORC1 (mammalian target of rapamycin complex 1) was found to blunt the chronic phase of hypertension, kidney injury, immune cell infiltration, renal hypertrophy, and cell proliferation in high‐salt (4.0% NaCl ) fed SS rats. Many of the upstream regulators of mTORC1 are known including amino acids, insulin/insulin‐like growth factor, and cellular energy. In SS rats, high‐salt intake is known to increase the renal production of reactive oxygen species (ROS) from the mitochondria and from NADPH oxidases (NOXs), especially NOX4 which is the most abundant NOX isoform in the kidney. NOX4 exclusively releases H2O2 and we recently reported that a chronic 3‐day renal interstitial infusion of H2O2 activates mTORC1 in vivo as evidenced by phosphorylation and activation of the ribosomal protein S6 (pS6S235/236/S6). In the present study, we hypothesized that activation of NOX4 and elevations of H2O2 in the high‐salt fed SS rat stimulate mTORC1 leading to renal injury. Western blot analysis found that mTORC1 activity was significantly higher in SS rats fed a high‐salt diet which also have high levels of H2O2 compared to SS‐NOX4 knock out rats (SSNox4−/−). SSNox4−/− rats with reduced H2O2 exhibited reduced level of mTORC1 activity (e.g., reduced phosphorylation of ribosomal protein S6 kinase 1, ribosomal protein S6) and mean arterial pressure (MAP) compared to SS rats fed a 4.0% NaCl diet for 21 days. It was also found in vitro that H2O2 activation of mTORC1 was direct and independent of phosphoinositide 3‐kinase (PI3K). Rapamycin treatment of SSNox4−/− rats fed a 4.0% NaCl diet for 21 days somewhat further reduced the already low levels of mTORC1 activity and immune cell infiltration (CD68, CD3), although no further reduction of MAP or renal injury was observed compared to vehicle treated SSNox4−/− rats. We conclude that H2O2 produced by NOX4 is a major upstream regulator of mTORC1 independent of PI3K activity and contributes importantly to blood pressure salt‐sensitivity and renal injury in SS rats.Support or Funding InformationHL‐116264