tbuckley/cabot-search huggingface dataset:
An E3 Ubiquitin Ligase-BAG Protein Module Controls Plant Innate Immunity and Broad-Spectrum Disease Resistance
Authors: Quanyuan You, Keran Zhai, Donglei Yang, Weibing Yang, Jingni Wu, Junzhong Liu, Wenbo Pan, Jianjun Wang, Xudong Zhu, Yikun Jian, Jiyun Liu, Yingying Zhang, Yiwen Deng, Qun Li, Yonggen Lou, Qi Xie, Zuhua He
Journal: Cell Host & Microbe (2016)
703. Development of RD-MolPack-trLNGFR/dCK.DM.S74E Stable Packaging Cells
Authors: Claudia Piovan, Erica Giuliani, Stefano Corna, Cinzia Scavullo, Sergio Bossi, Monika Pema, Jeffrey A. Medin, Monty McKillop, Bryan Au, Claudio Bordignon, Gian Paolo Rizzardi, Chiara Bovolenta
Journal: Molecular Therapy (2016)

The efficacy of several clinical approaches of adoptive T cell therapy (ACT) can be compromised by the insurgence of severe unfavorable events as graft-versus host disease (GvHD) secondary to bone marrow transplantation unless ex-vivo manipulation endow transplantable T cells with safety system. Introduction of drug-induced suicide genes, as for example the herpes simplex virus thymidine kinase (HSV-TK) transgene in T cells during bone marrow transplantation for leukemia and lymphoma treatment, represents a valid approach to control gene-modified cell proliferation and survival overtime. Transplanted modified cells are selectively deleted in vivo by specific activation of an otherwise inactive prodrug in those cells. Despite the well-documented utility of this approach, some limitations arose in terms of poor prodrug activation kinetics, escape from drug selection and immunological rejection.Engineered variants targeting the active site of the human deoxyCytidine Kinase (dCK) suicide enzyme were described as a powerful alternative to current strategies being able to activate a large variety of nucleoside analogue-based prodrugs, having low immunogenicity and permitting in vivo imaging of genetically modified cells. Lentiviral vector (LV)-mediated delivery of the triple mutant dCK.R104M.D133A.S74E (dCK.DM.S74E) fused to the truncated low density nerve growth factor receptor (LNGFR) selection marker renders human cells highly sensitive to the prodrugs bromovinyl-deoxyuridine (brivudine or BVdU), l-deoxythymidine (LdT), and l-deoxyuridine (LdU) via induction of apoptosis. MolMed has developed the proprietary RD-MolPack technology for the stable production of both second and third generation LVs, pseudotyped with the RD114-TR envelope that, in contrast to inducible systems based on VSV-G envelope, permits the generation of a simple, constitutive, no-toxic LV packaging system. We thus exploited the RD-MolPack technology to obtain several colonies of RD-MolPack-trLNGFR/dCK.DM.S74E producer cells by stable transfection of a SIN transfer vector plasmid carrying the trLNGFR/dCK.DM.S74E transgene followed by zeocin selection. The average titer calculated on CEM A3.01 target cells of several colonies is 1.9 × 105 ± 8.15 × 104 TU/106 cells/day, in line with the previous results obtained with the previously established RD2-MolPack-Chim3 and RD3-MolPack-GFP producer cells. These preliminary data suggest that the human suicide gene trLNGFR/dCK.DM.S74E can be constitutively produced in LV stable packaging cells without toxicity and its use can be therefore considered a valid option in the scenario of prodrug convertase enzymes.
477. ANGPTL4 Limits the Extent of Colonic Inflammation
Authors: Terri Phua, Dickson Shao Liang Chee, Nguan Soon Tan
Journal: Molecular Therapy (2016)

Gastrointestinal disorders such as inflammatory bowel disease, colitis and enteritis share signatures of an aggravated and protracted inflammatory response. As the first line of defense, epithelial cells play an important role in secreting chemoattractants and cytokines to launch or attenuate the immune response. However, little is known about the regulation of immune responses in the gastrointestinal tract. To understand the role of colonic epithelial cells during inflammation in C57BL/6J mice, we examine the effects of dietary C18 saturated (stearic acid), unsaturated (oleic acid) fat and dextran sulfate salt (DSS) in acute colonic inflammation. Microarray analysis was conducted comparing changes in global gene expression signatures between colon samples of three treatment groups. Preliminary studies suggest that the saturated diet resulted in a colonic gene expression profile that is more closely associated to that of DSS-treated. In addition, gene ontology analyses suggest that genes that overlap between all three treatment groups are mostly involved in immunological and metabolic diseases, as well as endocrine system disorders. Further interrogation revealed that an adipokine, angiopoietin-like 4 (ANGPTL4), modulates the degree and extent of colonic inflammation. We observed an elevated inflammation status, associated with massive immune cell infiltration, in ANGPTL4−/− mice compared to its ANGPTL4+/+ littermates during DSS-induced colonic inflammation. Our preliminary studies highlights the importance of regulating the colonic expression of ANGPTL4 as an avenue to attenuate gastrointestinal inflammation.
58. Engineered Cas9 Variants with Novel PAM Specificities Expand the Targeting Range of CRISPR/Cas Nucleases
Authors: Benjamin P. Kleinstiver, Michelle S. Prew, Ved V. Topkar, Shengdar Q. Tsai, Jae K. Joung
Journal: Molecular Therapy (2015)

Specificity and targeting range are important considerations when selecting an engineered nuclease platform for functional studies or therapeutic applications. The use of CRISPR/Cas9 to edit virtually any genome has enabled researchers to rapidly knock out genes or to introduce precise genomic changes. A number of studies have explored and improved the specificity of the CRISPR/Cas9 platform but the targeting range remains limited to certain sequences. The most widely used Cas9 from Streptococcus pyogenes (SpCas9) recognizes a protospacer adjacent motif (PAM) of the form NGG, a constraint that restricts the range of sequences that can be edited. Combined with the 5’ nucleotide requirement for guide-RNA expression from a polIII promoter, this restriction can be limiting particularly when one is using CRISPR/Cas9 nucleases to introduce precise mutations by homology-directed repair (HDR), which requires that double-strand breaks be introduced very closely to the site of sequence alteration. To address this challenge, we developed a heterologous genetic system that enabled us to rapidly screen libraries of Cas9 clones and identify variants with distinct cleavage specificities. Using this technology, we identified and optimized two different Cas9 variants that recognize novel PAMs. We demonstrate that these variants enable targeting of sites with non-canonical PAM sequences that were previously inaccessible with wild-type SpCas9, more than doubling the total range of sequences that can be edited by HDR and by error-prone non-homologous end-joining (NHEJ). We demonstrate that these variants can robustly modify endogenous genes in human cells and that they are compatible with previously described specificity improvements to SpCas9 such as truncated gRNAs. Furthermore, we also show Cas9 nucleases from other bacterial species such as Staphylococcus aureus and Streptococcus thermophilus can also be assessed for activity in our heterologous genetic system, suggesting that these alternative Cas9s might also be modified in their PAM specificities using the same approach we undertook with SpCas9. These orthogonal Cas9 nucleases offer the advantage of being substantially smaller in size than SpCas9, an important factor when considering packaging limits for viral delivery of CRISPR/Cas9 components. Taken together, our results more than double the targeting range of CRISPR/Cas9 nuclease platform and improve the prospects for translation of these reagents to clinical settings.
558. Second Generation Vaccinia Carrier for Scalable and Efficient Production of Recombinant AAV Vectors
Authors: Qizhao Wang, Zhongren Wu, Biao Dong, Weidong Xiao
Journal: Molecular Therapy (2015)

Recent successes of recombinant adeno-associated virus (rAAV) vectors in clinical trials demand a more efficient system for scalable production of high-quality rAAV vectors. Our recently developed rAAV production employed a novel concept by utilizing cytoplasmic carriers, vaccinia viruses carrying AAV rep and cap genes; nuclear carrier, an adenovirus carrier with AAV transgene expressing cassette and suspension Hela cells to produce rAAV vectors. In this prototype production system, two vaccinia virus carriers are necessary to provide the required Rep and Cap functions, with one containing Rep78 and VP2(Rep78-VP2) and another containing Rep52 and VP1(Rep52-VP1). This scheme is necessary due to the sequences identity between rep genes and cap genes. To further reduce the vaccinia carriers necessary for rAAV production, we developed a novel vaccinia carrier (VW22) which is capable of all necessary rAAV production gene products, Rep 78, Rep52, VP1, VP2 and VP3. Despite the VP1 and VP2, Rep78 and Rep52 all have identical amino acids in the C-termini, VW22 is stable for at least 12 continuous replication and production cycle. The rAAV packaging efficiency using VW22 is 2≈3 times higher than the previous generation using two vaccinia carriers expressing Rep78-VP2 and Rep52-VP1 separately. In the initial system, a wild type adenovirus was shown to increase rAAV yield significantly in the production cell line, HeLa cells. We made first improvement of the production system by making a HeLa cell line expressing adenovirus E1A and E1B (HE1 cells). HE1 cells were able to support rAAV replication efficiently without supplying wtAd. Higher rAAV yield in HE1 cells than that in Hela cells in the presence of wtAd were observed. Our new progresses reduced the numbers of vaccinia carrier and adenovirus helper. This new simplified system not only reduce the production cost, but also improved rAAV production yield and make the vector purification easier. We are able to further optimize other parameter for rAAV production such as the MOI of VW22 and Ad, the infection interval between Ad and VW22, the serum level and also the harvest time. In summary, the new development in using cytoplasmic carrier for packaging rAAV vectors showed great promise for a new system which is scalable, reliable, efficient and versatile for producing any serotype of AAV vectors.
1. Catecholamines and Hemodynamic Changes in Hypertension
Authors: Mitsuo Miyahara
Journal: Circulation (1966)

In order to evaluate a role of sympathetic nervous system in pathologic physiology of essential hypertension, an effect of autonomic ganglion blockade and noradrenaline on systemic and digital hemodynamics was observed in 55 normotensive subjects, 102 patients with essential hypertension and 18 patients with hypertension caused by glomerulonephritis (renal hypertension). Under basal condition, digital blood flow (DBF) was lowered, and digital (DVR) and total peripheral resistance (TPP) were elevated uniformly in both essential and renal hypertensives. Following the administration of hexamethonium, the increase of DBF and the decrease of DVR were significantly greater in essential hypertensives than in normotensives. These changes tended to be less remarkable in older individuals. Responses to noradrenaline were examined after the pretreatment of hexamethonium. Compared with the normotensive subjects, the elevation of DVR and TPR and the reduction of DBF was significantly greater, and the de-crease of cardiac index (CI) was observed in essential hypertensives. The majority of renal hypertensives showed the increase of DBF and CI, and no significant change in DVR and TPR. In contrast to noradrenaline, angiotensin 11 induced uniform changes of systemic and digital hemodynamics in normotensives and two hypertensive groups, i. e., the decrease of CI and DBF and the increase of DVR and TPR. Based on these findings, relationship between essential hypertension and sympathetic nervous tone was discussed. Coworkers; Dr. Osamu Iimura, Dr. Hisao Abe, Dr. Tsuyoshi Kikuiri, Dr. Tadashi Ueda, Dr. Ichiro Hirano. Dr. Takeshi Agata, Dr. Kenj Inami and Dr. Atushi Mukai.
Abstract SY36-03: Intraoperative molecular imaging with protease-activated fluorescent imaging agents
Authors: Melodi J. Whitley, Diana M. Cardona, Dan G. Blazer, Shelley Hwang, Rachel A. Greenup, Paul J. Mosca, Joan Cahill, Jeffrey K. Mito, Kyle C. Cuneo, Nicole Larrier, Erin O'Reilly, Ivan Spasojevic, Richard F. Riedel, William C. Eward, Linda G. Griffith, Moungi G. Bawendi, Jorge Ferrer, David B. Strasfeld, W. David Lee, Brian Brigman, David G. Kirsch
Journal: Cancer Research (2015)

Abstract Intra-operative detection of residual cancer in the tumor bed can be used to decrease the risk of a positive surgical margin, reduce the rate of re-excision, and tailor adjuvant therapy. LUM015 is a pegylated protease-activated imaging agent containing a near infrared fluorophore and quencher attached by a polypeptide linker. Using mouse models of soft tissue sarcoma (STS), we showed that LUM015 selectively localizes to the tumor and upon cleavage of the linker by proteases in the tissue, the quencher is released, allowing fluorescence to be detected. We recently completed a phase I clinical trial to test the safety of LUM015 in human patients with cancer. This open-label nonrandomized trial compared 3 dose cohorts (0.5, 1.0, and 1.5 mg/kg) of LUM015 in order to determine a safe dose of LUM015 that labels tumors in humans. A total of 15 subjects, 12 with STS and 3 with breast cancer, received IV LUM015 prior to surgical resection without any adverse pharmacological activity (APA). Quantitative fluorescence imaging of the resected tissues revealed that tumor fluorescence was significantly higher than corresponding normal tissue from the same patient (p&amp;lt;0.0001) with a mean tumor to normal fluorescence ratio of 5.4. Furthermore, within the study population, the distribution of tumor fluorescence values was significantly higher than those recorded for fat or muscle (p&amp;lt;0.009). We conducted comparative analyses of LUM015 pharmacodynamics in mouse and human subjects showing that biodistribution into tumors and activation by proteases is conserved across species. These studies revealed the presence of a novel, small molecule metabolite that correlates strongly with tissue fluorescence. Additional in vivo studies in mice and in vitro studies with mouse and human tissues showed that LUM015 is selectively distributed to and accumulates in tumors to result in increased fluorescence when compared to normal tissues. Citation Format: Melodi J. Whitley, Diana M. Cardona, Dan G. Blazer, Shelley Hwang, Rachel A. Greenup, Paul J. Mosca, Joan Cahill, Jeffrey K. Mito, Kyle C. Cuneo, Nicole Larrier, Erin O'Reilly, Ivan Spasojevic, Richard F. Riedel, William C. Eward, Linda G. Griffith, Moungi G. Bawendi, Jorge Ferrer, David B. Strasfeld, W. David Lee, Brian Brigman, David G. Kirsch. Intraoperative molecular imaging with protease-activated fluorescent imaging agents. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr SY36-03. doi:10.1158/1538-7445.AM2015-SY36-03
A novel BCMA/CD3 bispecific T-cell engager for the treatment of multiple myeloma induces selective lysis in vitro and in vivo
Authors: S Hipp, Y-T Tai, D Blanset, P Deegen, J Wahl, O Thomas, B Rattel, P J Adam, K C Anderson, M Friedrich
Journal: Leukemia (2016)

B-cell maturation antigen (BCMA) is a highly plasma cell-selective protein that is expressed on malignant plasma cells of multiple myeloma (MM) patients and therefore is an ideal target for T-cell redirecting therapies. We developed a bispecific T-cell engager (BiTE) targeting BCMA and CD3ɛ (BI 836909) and studied its therapeutic impacts on MM. BI 836909 induced selective lysis of BCMA-positive MM cells, activation of T cells, release of cytokines and T-cell proliferation; whereas BCMA-negative cells were not affected. Activity of BI 836909 was not influenced by the presence of bone marrow stromal cells, soluble BCMA or a proliferation-inducing ligand (APRIL). In ex vivo assays, BI 836909 induced potent autologous MM cell lysis in both, newly diagnosed and relapsed/refractory patient samples. In mouse xenograft studies, BI 836909 induced tumor cell depletion in a subcutaneous NCI-H929 xenograft model and prolonged survival in an orthotopic L-363 xenograft model. In a cynomolgus monkey study, administration of BI 836909 led to depletion of BCMA-positive plasma cells in the bone marrow. Taken together, these results show that BI 836909 is a highly potent and efficacious approach to selectively deplete BCMA-positive MM cells and represents a novel immunotherapeutic for the treatment of MM.
TT30, a Novel Regulator of the Complement Alternative Pathway (CAP), Inhibits Hemolysis of Paroxysmal Nocturnal Hemoglobinuria (PNH) Erythrocytes and Prevents Upstream C3 Binding On Their Surface in An in Vitro Model.
Authors: Antonio M. Risitano, V. Michael Holers, Bruno Rotoli
Journal: Blood (2009)
Relative Trends in Myocardial T2* Versus Myocardial Iron Concentration As Representations of Myocardial Iron Chelation Efficacy in Patients with β Thalassemia Major Treated with Deferasirox for up to 3 Years
Authors: John B Porter, Ali T Taher, Yesim Aydinok, Amal El-Beshlawy, Mohsen Elalfy, Ali El-Ali, Nathalie Barbier, M Domenica Cappellini
Journal: Blood (2014)
SURG-10. IMPACT OF INSURANCE STATUS AND RACE ON RECEIPT OF SURGERY FOR ACOUSTIC NEUROMA: A NATIONAL CANCER DATABASE ANALYSIS
Authors: Shearwood McClelland, James D Murphy, Charles R Thomas
Journal: Neuro-Oncology (2016)

The traditional management of acoustic neuroma (AN) has involved microsurgical excision. Previous inpatient studies have demonstrated that in-hospital morbidity/mortality following AN surgery differs by patient race and insurance status; however there have been no studies addressing the impact of these demographics on the receipt of surgery on a nationwide level. The National Cancer Data Base (NCDB) from 2004-2013 was used to identify patients who received surgical resection as the sole treatment for AN. A multivariable logistic regression model characterized patients who received surgery, adjusting for patient age, sex, income, medical comorbidities, and tumor size. 10,136 patients received AN surgery. Patients who were African-American (OR=0.7;95%CI=0.5-0.9;p=0.01), at least age 65 (OR=0.4;95%CI=0.4-0.6;p<0.0001), had Medicare insurance (OR=0.6;95%CI=0.4-0.7;p=0.0005), or treated at a community hospital (OR=0.4;95%CI=0.2-0.7;p=0.007) were less likely to receive surgery. Patients who had Medicaid insurance (OR=1.2;95%CI=0.8-1.8;p=0.04), a Charlson/Deyo score of one (OR=1.9;95%CI=1.5-2.3;p=0.002), residence in the central United States (OR=1.6;95%CI=1.4-1.9;p=0.006), or treated at an integrated network (OR=1.2;95%CI=0.9-1.6;p=0.0004) were more likely to receive surgery. Elderly age, African-American race (by 30%), Medicare insurance (by 40%) and treatment at a community hospital (by 60%) were independent predictors for reduced receipt of AN surgery. Conversely, Medicaid insurance (by 20%), Charlson/Deyo score of one, central United States location, and treatment at an integrated network (by 20%) independently predicted increased receipt of AN surgery. Given that the adjusted relative risk for in-hospital mortality of AN surgery for African-Americans is 10.6 compared with Caucasians (Curry et al., 2009), and that private insurance reduces in-hospital and perioperative morbidity by 42% compared with non-private insurance (McClelland et al., 2011), these findings indicate that while it may be beneficial for African-American and Medicare insurance patients to be steered away from surgery, it is likely detrimental for Medicaid insurance patients to be steered towards AN surgery.
In chronic severe functional constipation, electroacupuncture increased complete spontaneous bowel movements
Authors: Subhankar Chakraborty, Adil E. Bharucha
Journal: Annals of Internal Medicine (2016)

ACP Journal Club20 December 2016In chronic severe functional constipation, electroacupuncture increased complete spontaneous bowel movementsSubhankar Chakraborty, MD, PhD, Adil E. Bharucha, MBBS, MDSubhankar Chakraborty, MD, PhDMayo Clinic, Rochester, Minnesota, USA (S.C., A.E.B.)Search for more papers by this author, Adil E. Bharucha, MBBS, MDMayo Clinic, Rochester, Minnesota, USA (S.C., A.E.B.)Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/ACPJC-2016-165-12-069 SectionsAboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail Source CitationLiu Z, Yan S, Wu J, et al. Acupuncture for chronic severe functional constipation: a randomized, controlled trial. Ann Intern Med. 2016 Sep 13. [Epub ahead of print]. https://pubmed.ncbi.nlm.nih.gov/27618593Clinical Impact RatingsGIM/FP/GP: Gastroenterology: References1 American Gastroenterological Association, Bharucha AE, Dorn SD, Lembo A, Pressman A. Gastroenterology. 2013;144:211-7. [PMID: 23261064] Google Scholar2 Bharucha AE, Pemberton JH, Locke GR 3rd. Gastroenterology. 2013;144:218-38. [PMID: 23261065] Google Scholar3 Jin X, Ding YJ, Wang LL, et al. Zhongguo Zhen Jiu. 2010;30:97-101. [PMID: 20214063] Google Scholar4 Wang LJ, Wang LL. Zhongguo Zhen Jiu. 2011;31:320-4. [PMID: 21528597] Google Scholar Author, Article, and Disclosure InformationAffiliations: Mayo Clinic, Rochester, Minnesota, USA (S.C., A.E.B.)This article was published at Annals.org on 6 December 2016. PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics Cited byElectroacupuncture pretreatment attenuates brain injury in a mouse model of cardiac arrest and cardiopulmonary resuscitation via the AKT/eNOS pathwayAcupuncture in Inflammatory Bowel Disease 20 December 2016Volume 165, Issue 12Page: JC69KeywordsAcupunctureAdverse eventsConstipationHeart diseasesIntent to treat analysisMedicareNumber needed to treatPacemakersQuality of lifeQuestionnaires ePublished: 20 December 2016 Issue Published: 20 December 2016 Copyright & PermissionsCopyright © 2016 by American College of Physicians. All Rights Reserved.PDF downloadLoading ...