tbuckley/cabot-search huggingface dataset:
Abstract 5302: Control of Reversible Phosphorylation of Ryanodine Receptor by <i>MiR-1</i> : Mechanism of Cardiac Arrhythmia
Authors: Dmitry Terentyev, Andriy Belevych, Radmila Terentyeva, Donald E Kuhn, Geraldine E Malana, Mickey M Martin, Maha Abdellatif, David S Feldman, Terry S Elton, Sandor Gyorke
Journal: Circulation (2008)

MicroRNAs are small endogenous noncoding RNAs that regulate protein expression by hybridization to imprecise complementary sequences of target mRNAs. Changes in abundance of muscle-specific microRNA, miR-1 , have been implicated in cardiac disease, including arrhythmia and heart failure. However, the specific molecular targets and cellular mechanisms involved in the action of miR-1 in the heart are only beginning to emerge. In this study we investigated the effects of increased expression of miR-1 on excitation-contraction coupling and Ca cycling in adult rat ventricular myocytes by using methods of electrophysiology, confocal Ca imaging and quantitative immunoblotting. Adenoviral-mediated overexpressions of miR-1 in myocytes resulted in a marked increase in the amplitude of the inward Ca current and flatten cytosolic Ca transients voltage dependency. The frequency of spontaneous Ca sparks recorded in intact resting cells was enhanced, while the sarcoplasmic reticulum Ca content was reduced in miR-1 - overexpressing myocytes as compared with controls. In the presence of beta-adrenergic receptor agonist isoproterenol, rhythmically paced miR-1 -overexpressing myocytes exhibited spontaneous arrhythmogenic oscillations of intracellular Ca, events that occurred only rarely in control myocytes. The effects of miR-1 were completely reversed by the CaMKII inhibitor KN93. Immunological analysis with phospho-specific antibodies showed that while phosphorylation of phospholamban was not altered, miR-1 overexpression increased phosphorylation of the ryanodine receptor at Ser-2814 (CaMKII) but not at Ser-2808 (PKA). Overexpression of miR-1 was accompanied by a selective decrease in expression of the protein phosphatase PP2A regulatory subunit B56alpha involved in PP2A targeting to specialized subcellular domains. We conclude that miR-1 through translational inhibition of this mRNA target, causes CaMKII-dependent hyperphosphorylation of RyR2 via disrupting localization of PP2A activity to this channel, enhances RyR2 activity, and promotes arrhythmogenic SR Ca release. This mechanism could contribute to induction of arrhythmia in disease states accompanied by elevated miR-1 . This research has received full or partial funding support from the American Heart Association, AHA National Center.
Abstract 18593: Use of a CRISPR/Cas System for Cardiovascular Disease Modeling and Therapeutic Applications
Authors: Qiurong Ding, Chad A Cowan, Kiran Musunuru
Journal: Circulation (2013)

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have recently emerged as a new technology with which to perform genome editing in mammalian cells. We have tested the targeting efficiency of a CRISPR/Cas system in human pluripotent stem cells (hPSCs) by designing CRISPRs targeting eight loci in seven genes--AKT2, CELSR2, CIITA, GLUT4, LDLR, LINC00116, and SORT1. We found that different CRISPRs yield mutant clones at efficiencies of 51%-79%, with homozygous mutant clones generated at efficiencies of 7%-25%--far superior to any other genome-editing tool evaluated to date. Using this CRISPR/Cas system, we have generated isogenic cell lines with and without disease mutations, followed by differentiation into different cell types relevant to cardiovascular and metabolic diseases, including hepatocytes, adipocytes, and cardiomyocytes, which allows for rigorous human disease modeling and elucidation of new disease mechanisms. For example, using this system we have discovered that the novel gene LINC00116 has a role in lipid metabolism in adipocytes. We are building on these initial results by testing the CRISPR/Cas system for in vivo genome editing in mice, with a view towards establishing a novel therapeutic modality. We have delivered in vitro transcribed RNAs encoding Cas9 nuclease and CRISPRs targeting genes of therapeutic interest--e.g., the PCSK9 gene--formulated in lipid nanoparticles specific to mouse liver, with the intent of knocking out the gene in liver. These experiments provide a proof-of-principle that a CRISPR/Cas system could be used for human therapeutic applications in the future.
A prospective phase II trial of erlotinib in advanced non-small cell lung cancer (NSCLC) patients (p) with mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR)
Authors: L. Paz-Ares, J. M. Sanchez, A. García-Velasco, B. Massuti, G. López-Vivanco, M. Provencio, A. Montes, D. Isla, M. L. Amador, R. Rosell
Journal: Journal of Clinical Oncology (2006)

7020 Background: The EGFR TK inhibitor erlotinib is a standard treatment in previously treated NSCLC p. Recently, EGFR gene mutations have been reported to be a predictor of efficacy of this and related compounds. This phase II trial evaluated the efficacy and safety of erlotinib in untreated p with advanced NSCLC and mutated EGFR. Methods: Eligibility criteria included stage IIIB-IV chemotherapy-naive p with confirmed NSCLC and mutated EGFR, PS 0–2, age ≥ 18 years, adequate organ functions, measurable disease and written informed consent. DNA was extracted by laser capture microdissection. Exon 19 deletions were examined by Genscan and the L858R and T790M mutations with a TaqMan assay. Mutations were confirmed by sequencing. Erlotinib was given orally at 150 mg per day. Results: From March to December 2005, 37 (12.5%) of 297 tumors screened were found to have mutations in the TK domain of the EGFR gene (25 deletions; 11 L858R; no T790M). Median age: 68 years; 12 males, 25 females; histology: 28 adenocarcinoma, 4 BAC, 4 large cell carcinoma; 9 smokers. Response has been evaluated in 21 p so far: complete response, 6 p; partial response, 13 p; progressive disease, 2 p; overall response rate, 90% (95% CI:70%-90%). Responses occurred in 100% of tumors with exon 19 deletions and in 75% of those with L858R. All responses are ongoing (up to ≥8 months) and all but one p are still alive. Toxicity profile was predictable (mild to moderate rash, diarrhea, asthenia and emesis). No drug related severe adverse events occurred. Conclusions: Single-agent erlotinib produces a high response rate in previously untreated p with advanced NSCLC and mutated EGFR. In-frame deletions in exon 19 seem particularly predictive for response in this setting. [Table: see text]
Abstract 5423: rAAV-Mediated Gene Therapy with Inducible Nitric Oxide Synthase (iNOS) Affords Permanent Cardioprotection (1 Year) without Adverse Functional Consequences
Authors: Qianhong Li, Yiru Guo, Wen-Jian Wu, Qinghui Ou, Santosh K Sanganalmath, Wei Tan, Xiaoping Zhu, Jingyao Mu, Buddhadeb Dawn, Roberto Bolli
Journal: Circulation (2008)

The ultimate goal of prophylactic gene therapy is to confer permanent protection against ischemia. Although gene therapy with iNOS is known to protect against myocardial infarction at 3 days and up to 2 months, the long-term effects of iNOS gene therapy on myocardial ischemic injury and function are unknown. To address this issue, we created a recombinant adeno-associated viral vector carrying the iNOS gene (rAAV/iNOS) which enables long-lasting transgene expression. Mice received injections in anterior LV wall of rAAV/LacZ or rAAV/iNOS; 1 year later, they underwent a 30-min coronary occlusion (O) and 4 h of reperfusion (R). iNOS gene transfer resulted in elevated iNOS protein expression (+ 2.9-fold vs. LacZ group, n=6, P&lt;0.05; Fig ) and iNOS activity (+ 3.3-fold vs. LacZ group, n=4, P&lt;0.05) 1 year later. Infarct size (% of risk region) was dramatically reduced at 1 year after iNOS gene transfer (13.5+/−2.2%, n=12, vs. 42.9+/−2.6%, n=12, in LacZ group; Fig ). The infarct-sparing effects of iNOS gene therapy at 1 year were as powerful as those observed 24 h after ischemic PC (six 4-min O/4-min R cycles) (16.3+/−2.3%, n=8; Fig ). Importantly, compared with the LacZ group (n=11), iNOS gene transfer (n=10) had no effect on LV dimensions or function for up to 1 year (at 1 year: LVEDD 4.4+/−0.1 vs. 4.2+/−0.2 mm; LVESD 2.9+/−0.1 vs. 2.9+/−0.2 mm; FS 34+/−1.8 vs. 32+/−2.6%; EF 56+/−2.3 vs. 60+/−2.9%) (echocardiography). These data demonstrate, for the first time, that rAAV-mediated iNOS gene transfer affords long-term, probably permanent (1 year) cardioprotection without adverse functional consequences, providing a strong rationale for further preclinical testing of prophylactic gene therapy.
Integrating people who are stigmatized: the tetralogue model.
Authors: Petr Nawka, Charlene M Reiss
Journal: World Psychiatry (2002)

Although present everywhere, segregation of people with mental illness and their relatives and violation of their human rights cannot be justified any longer. What can change the status quo? Breaking the silence of a society that does not recognize or acknowledge its mental health problems. Communicating about them opens the doors for the integration of mental health into general health and people with mental illness into society. In Michalovce, Slovakia, the tetralogue model (a dialogue of four sides) supports this necessary communication to overcome stigma by integrating people with mental illness into society, bringing benefit to all parties involved: people with mental illness, their relatives, professionals and the public.

First and foremost, we recognized that people with mental illness and their relatives are the best experts on issues of stigma and discrimination. The motivation to improve their own situation needs to be supported. Having formed associations in 1997, users and relatives are able to act as equal partners with the professionals, moving from dialogue to trialogue.

The first result of regular monthly meetings of the trialogue was a patients' advocacy programme, changing the way that patients and professionals relate to each other. Each week, individuals from the local users' organization visit the hospital, hearing the concerns of patients and finding solutions by working as partners with the professional staff. At first, the nurses expressed resistance to this programme, fearing criticism and rebellion. In a short time, they found that addressing problems openly helped solve them more quickly, with less stress and conflict. In addition, patients have started to contribute to their own treatment plans. The professionals are finding their work easier as they share the responsibility for treatment with the patients, rather than taking the traditional, patriarchal, non-participatory approach.

Real integration is not possible without winning over the public. People with mental illness must be the main actors who, together with their relatives and professionals, help the public overcome fear and anger. Our Club of People with Schizophrenia is talking to school groups and meeting with public administrators. The response from the participants - the presenters and the audiences, especially the students - has been overwhelmingly positive. We have seen these interactions benefit society by encouraging individuals to become more interested in their own mental health.

Our participation in the WPA Program against Stigma and Discrimination has enabled us to involve more people across Slovakia in our work. This not only strengthens the Slovak users' and relatives' movement, but also motivates professionals to participate in this partnership. The result is a stronger tetralogue at the national level. As more people at all levels become interested in mental health, the walls around society can slowly come down, replaced with bridges to mental health and open doors to integration.
Abstract 1632: Coronary Calcification Relates to Bone Metabolism and CD4+ T-Cell Activation
Authors: Hirofumi Soejima, Takashi Fukunaga, Yoko Oe, Ryuichiro Fukushima, Hisao Ogawa
Journal: Circulation (2007)

Background: Osteopontin (OPN) facilitates bone resorption by enhancing the binding of osteoclast to hydroxyapatite. OPN is expressed in both the intima and the medial layers when arterial calcification is present. OPN is the most abundant molecule secreted by activated T-cells. We previously demonstrated that CD4+ T-cells play an important role in acute coronary syndrome and coronary artery disease (CAD). Osteoprotegerin (OPG) is associated with the progression of vascular calcification in diabetic patients or in patients on long term hemodialysis. Either an intravenous injection of recombinant OPG protein or the transgenic overexpression of OPG has been shown to reverse osteoporosis and prevent vascular calcification. Collagen Type I constitutes the most substantive proportion of the matrix protein of bone. Analysis of a Collagen Type I breakdown product, a Cross-linked N-terminal Telopeptide of Type I Collagen (NTX), has been used to estimate rates of bone degradation. We therefore investigated whether these markers (OPN, OPG, NTX) reflected coronary calcification as detected by multislice computed tomography (MSCT) in patients with CAD and assessed the association these markers with CD4+ T-cell activation. Methods and Results: We measured serum OPN, OPG, and NTX levels of 76 consecutive patients who received MSCT. The frequency of OPN producing CD4+ T-cells was determined with flow cytometry. Coronary calcification was present in 41 of the 76 patients. Serum OPN, OPG, and NTX levels were all significantly higher in patients with coronary calcification than in those without. Multiple logistic regression analysis revealed that age &gt;70 years, hypertension and serum OPN level &gt;630 ng/ml were determinants of coronary calcification independent of other coronary risk factors. The frequency of osteopontin producing CD4+ T-cells was higher in patients with coronary calcification and it was negatively correlated with serum OPG levels. Conclusions: The presence of coronary calcification is reflected better by an osteoporosis facilitating factor (OPN) than by an osteoporosis inhibiting factor (OPG) or by a marker of the bone degradation rate (NTX). Assessment of bone metabolism could be useful for clinical determinations of coronary heart disease.
Abstract 15265: Extent of Elevation in Omega-3 Polyunsaturated Fatty Acid Levels in Blood is Directly and Strongly Related to Increases in Atrial Refractoriness and Susceptibility to Inducible Atrial Fibrillation in Humans
Authors: Saurabh Kumar, Fiona Sutherland, Paul B Sparks
Journal: Circulation (2011)

Background: The relationship between omega-3 polyunsaturated fatty acid (n-3 PUFA) intake and risk of atrial fibrillation (AF) has been inconsistent in epidemiological studies. Few studies have use...
Abstract 5690: Idiopathic Catecholamine-sensitive Epicardial Ventricular Tachycardia At The Crux Of The Heart
Authors: Harish Doppalapudi, Karthik Ramaswamy, Joon Ahn, Takumi Yamada, G Neal Kay
Journal: Circulation (2008)

Idiopathic ventricular tachycardia (VT) has been described from the epicardial surface of the left ventricle (LV), usually near the summit of the LV in the outflow tract. Ablation of these VTs may be possible by delivery of energy within the coronary venous system or directly on the epicardial surface. We describe a distinct syndrome of focal epicardial VT induced by catecholamine infusion that arises from the crux of the heart. Among 340 cases of idiopathic VT referred for catheter ablation, 4 patients were identified with a clinical syndrome of catecholamine sensitive VT that was mapped to the epicardial surface at the crux of the heart. There were 3 males and 1 female (age 31–79 yrs, mean 58). VT was sustained in all patients and associated with syncope or near syncope in 3 of 4 pts. The LVEF was &gt;0.55 in 3 pts and mildly depressed (0.45) in 1 pt. In all pts VT could be induced with programmed stimulation or burst pacing from the right ventricular apex but required the infusion of isoproterenol for induction in 3. The VT was very rapid with a mean cycle length of 264 msec. The surface ECG during VT demonstrated a left superior axis QRS morphology in all pts, with an abrupt precordial tansition from V1 to V2 in 3 pts and R waves across the precordium in one. The precordial maximal deflection index was &gt; 0.55 in all pts (mean 0.67). The site of earliest activation during intracardiac mapping occurred at the crux of the heart with activation in the middle cardiac vein or proximal coronary sinus recorded 20 –50 msec (mean −38 msec) prior to the onset of the surface QRS. Catheter ablation with irrigated RF was attempted within the middle cardiac vein or proximal coronary sinus in all pts and was successful in 1. In 2 of 3 remaining pts, percutaneous epicardial RF ablation was attempted and was successful. Simultaneous coronary angiography demonstrated the site of earliest activation within 5–10 mm of the proximal posterior descending coronary artery (PDA). There was no acute narrowing of the PDA in any pt following ablation. Idiopathic VT may arise from the epicardial surface at the crux of the heart in close proximity to the PDA. This syndrome can result in very rapid, catecholamine sensitive VT, and may require careful attention to the PDA during ablation.
Abstract 17318: Caspase Inhibition Prevents Hypertrophy and Improves Cardiac Function with Chronic Pressure Overload
Authors: Misun Park, Jung Ah Lee, Shumin Gao, SeongHun Yoon, Lai-Hua Xie, Ludny Charles, Samuel J Leibovich, Richard N Kitsis, Stephen F. Vatner, Dorothy E. Vatner
Journal: Circulation (2011)

It is widely held that a major mechanism underlying the transition from stable left ventricular (LV) function to failure is myocyte loss through apoptosis. To test this hypothesis, we examined the effects of chronic transverse aortic constriction (TAC) in C57BL/6 mice with or without administration of the caspase inhibitor (CI) (Z-Asp-2,6-DCBMk) delivered through an intraperitoneal osmotic minipump. In mice treated with CI, LV function (LV ejection fraction) was preserved (73±1%) compared with the vehicle group (52±7%) (p&lt;0.05) after 2 weeks of TAC. One possibility was that CI prevented apoptosis of myocytes and thereby improved LV function. This was not likely, since with CI, apoptosis was reduced, but relatively trivially in myocytes; using double staining with TUNEL and wheat germ agglutinin, only 7% of the reduced cellular apoptosis in the heart could be attributed to myocytes as compared with 93% of non-myocytes protected. Furthermore, if myocyte apoptosis protection was the mechanism, then LV mass would likely have been larger with CI. However, the reverse occurred, i.e., CI actually prevented LV hypertrophy; LV weight/tibial length did not increase significantly with CI after TAC (5.26±0.30) vs. sham (4.95±0.30) as opposed to vehicle after TAC (7.49±0.43). This was not because of differences in LV/aortic pressure gradient in CI (102±4mmHg) vs. vehicle (100±2mmHg), but likely due to protected LV wall stress, which was less (p&lt;0.05) in the CI group (70.2±9.9 Kdyn/cm 2 ) than vehicle (132.2±19.4 Kdyn/cm 2 ). Therefore, the mechanism for improved function with TAC and CI was likely related to an effect of CI on myocytes. Accordingly, we examined isolated myocytes using a Langendorff perfusion apparatus, and measured isolated myocyte contractility by an optical edge-tracking method. After TAC the contractility (% of cell length) under 1mM calcium was increased in CI (15.5±1.2%) compared to vehicle (10.2±1.4%) (p&lt;0.01). Thus, CI improved LV function after chronic pressure overload, not by protecting against myocyte apoptosis, but rather by improving myocyte contractile function resulting in reduced LV wall stress and hypertrophy.
Abstract 5757: Magnetic Resonance Imaging with Iron-Oxide Labeling Detects Differential Cell Survival after Doxorubicin Exposure in Cardiac Myocytes, Fibroblasts, and Stem Cells
Authors: Rajesh Dash, Trevor Chan, Mayumi Yamada, Marietta Paningbatan, Phillip M Swigart, Bat-Erdene Myagmar, Phillip C Yang, Paul C Simpson
Journal: Circulation (2008)

Heart failure caused by anthracycline chemotherapy, e.g. doxorubicin (DOX), is preceded by significant cell apoptosis. Early, non-invasive detection of cardiac cell death may improve patient outcomes by enabling real-time adjustment of chemotherapy. To this end, Annexin V protein (ANX) binds to membrane-bound phosphatidylserine, which is externalized in early apoptosis. We tested whether ANX conjugated to superparamagnetic iron oxide (SPIO) could detect and quantify apoptosis via T2-weighted magnetic resonance imaging (MRI). ANX was purified and conjugated to SPIO. Cultured, neonatal rat cardiac myocytes (CMs) and fibroblasts (FBs), and mouse mesenchymal stem cells (mMSCs) were exposed to DOX (1uM) for varying times, then stained with ANX-SPIO, or with fluorescent ANX (ANX-FLUOS, Roche) or TUNEL as controls. In vitro MRI (3 Tesla, GE Excite, WI) was performed on plated cells using a gradient echo (GRE) sequence (see Table ). MRI signal after ANX-SPIO incubation correlated highly with the number of ANX-FLUOS or TUNEL positive cells after exposure to varying durations of DOX (R=0.91 ANX-FLUOS vs. ANX-SPIO, n=5; R=0.80 TUNEL vs. ANX-SPIO, n=4; p&lt;0.05). ANX-SPIO signal was detectable by MRI from as few as 15–20 apoptotic mMSCs. CMs exhibited the highest sensitivity to DOX (see Table ), with peak apoptosis after 30 minutes of DOX exposure. Conversely, cardiac FBs were resistant to DOX for 22 hours, and mMSCs showed intermediate susceptibility to DOX. Competition binding between ANX-SPIO and ANX-FLUOS confirmed the specificity of ANX-SPIO for apoptotic cells in culture. In vitro MRI of a magnetically-labeled marker of apoptosis (ANX-SPIO) can accurately detect cell death in culture with high sensitivity. MRI of ANX-SPIO can also resolve differential susceptibility to DOX in CMs, FBs, and mMSCs. This compound may hold promise for in vivo MRI detection of early anthracycline-induced cell death in the myocardium. This research has received full or partial funding support from the American Heart Association, AHA Western States Affiliate (California, Nevada &amp; Utah). Cell-Type Specific Susceptibility to Doxorubicin Assessed by MRI