A New Therapeutic Concept Based on Hyperstabilization of Sense-Antisense Duplexes.

We proposed that known helix-stabilizing, minor-groove binding agents such as CC-1065 and its analog, U-71,184, might be feasibly targeted to specific mRNA by tethering them to antisense oligonucleotides. The CC-1065-tethered antisense oligonucleotide might be hyperstabilized to the target mRNA and arrest the translation of mRNA. Our hypothesis is that these agents can bind and increase the stability of RNA-DNA duplexes which may occur between mRNA and antisense oligonucleotides. To verify our hypothesis, we evaluated the ability of CC-1065, U-71,184, and distamycin A to bind and hyperstabilize the 20-mer sense sequence (5\sp\prime-TTACTTCAGTTATGAGACCA-3\sp\prime) duplexed with complementary (antisense) oligonucleotide sequence made with phosphodiester (PO), phosphorothioate (PS), or methyl phosphonate (MP) linkages. Sense DNA sequences (20 mer), when duplexed with PO, PS, or MP DNA oligonucleotides, were saturated at 2, 2, and 1 CC-1065 molecules per duplex, respectively, or at 1 molecule of U-71,184 for each duplex. Each molecule of CC-1065 and U-71,184 was able to bind to poly(rA)-oligo(dT)\sb{20} at maximum binding. Similarly, a sense RNA sequence duplexed with either PO or PS structures was saturated at one CC-1065 molecule per duplex. It was apparent from the melting temperature study that CC-1065 was generally more effective than U-71,184 against DNA-DNA duplexes even if the antisense DNA strand has modified backbone structures. Only CC-1065 was able to hyperstabilize the RNA-DNA duplex as evidenced by a 29\sp\circC increase of the melting temperature. U-71,184 and distamycin A were able to increase the melting temperature of RNA-DNA duplex only about 2\sp\circC and 4\sp\circC, respectively. Translation (wheat germ extract) was inhibited in a dose-dependent manner at antisense:mRNA ratios ranging from 10 to 72. Concomitant incubations of the duplexes with CC-1065 caused significantly enhanced depression of translation at the higher doses. CC-1065 appears to induce hyperstabilization between the antisense oligonucleotide and the mRNA target and arrest translation. Further development of hyperstabilizing antisense oligonucleotide structures is amply justified

In Vitro Chemosensitivity Using the Histoculture Drug Response Assay in Human Epithelial Ovarian Cancer

The choice of chemotherapeutic drugs to treat patients with epithelial ovarian cancer has not depended on individual patient characteristics. We have investigated the correlation between in vitro chemosensitivity, as determined by the histoculture drug response assay (HDRA), and clinical responses in epithelial ovarian cancer. Fresh tissue samples were obtained from 79 patients with epithelial ovarian cancer. The sensitivity of these samples to 11 chemotherapeutic agents was tested using the HDRA method according to established methods, and we analyzed the results retrospectively. HDRA showed that they were more chemosensitive to carboplatin, topotecan and belotecan, with inhibition rates of 49.2%, 44.7%, and 39.7%, respectively, than to cisplatin, the traditional drug of choice in epithelial ovarian cancer. Among the 37 patients with FIGO stage Ⅲ/Ⅳ serous adenocarcinoma who were receiving carboplatin combined with paclitaxel, those with carboplatin-sensitive samples on HDRA had a significantly longer median disease-free interval than patients with carboplatin- resistant samples (23.2 vs. 13.8 months, p＜0.05), but median overall survival did not differ significantly (60.4 vs. 37.3 months, p＝0.621). In conclusion, this study indicates that HDRA could provide useful information for designing individual treatment strategies in patients with epithelial ovarian cancer

A Design and Development of the Learning Contents Management based on the Personalized Online Learning

Teaching-learning methods are undergoing rapid transformation in terms of new information and communication technology and in accordance with onset of the 4th Industrial Revolution. The educational environment is being transformed into various forms, with examples being found not only in the existing traditional educational environment, but also in online education and blended learning. Existing online learning (LMS, LCMS) is offered in a limited contents transmission online educational environment, and has been limited but the level of support offered to a learner’s personalized learning. This study will overview existing flexible model of contents, suggest possible problems, and attempt to solve these problems. LCMS was designed and realized based on the open source Moodle platform, offering personalized contents to learners. LCMS is composed of the following 3 functions: contents registration of metadata inputted by administrator; search functionality for personalized learner contents; and personalized contents automatically being recommended to learners. As a result of the research, we made online learning environment that can provide customized learning recommendation and self - directed learning by increasing the continuity and efficiency of learning by automatically providing customized online contents to learners. Through this study, the learning of students promises to be effectively initiated by being based on available LCMS functions related to personalized educational contents in online education

Value of Professional Sport Teams in the Community: Reexamining the Measurement Properties of the Community Impact Scale

The Community Impact Scale was developed by Zhang et al. (1996) to measure essential value dimensions of professional sports in the community. The original scale includes 45 items under eight factors (Community Solidarity, Public Behavior, Social Equity, Pastime Ecstasy, Health Awareness, Individual Quality, Excellence Pursuit, and Business Opportunity). To further ensure its usefulness for theoretical and practical applications, this study re-examined measurement properties of the scale through conducting a confirmatory factor analysis (CFA). Research participants (N = 349) were residents of three major metropolitan areas, who responded to a survey in various community settings. With some modifications and eliminations of items, the CFA revealed that the eight-factor model with 32 items provided a good fit to the data (e.g., RMSEA = .060; SRMR = .053; CFI =.910). The resolved scale also displayed good convergent and discriminant validity, as well as internal consistency. Discussions are made with respect to the merits of the scale and its research applications in addressing social, political, marketing, and financing issues of professional sport teams

Comparative effects of norepinephrine and vasopressin on internal thoracic arterial graft flow after off-pump coronary artery bypass grafting

ObjectiveVasoconstrictors such as norepinephrine and vasopressin are commonly used to raise the blood pressure during myocardial revascularization. The internal thoracic artery is commonly used for coronary artery grafting because of its long-term patency. However, the internal thoracic artery is a living conduit that responds to vasoactive substances. The objective of this study was to measure change in internal thoracic arterial flow after infusion of norepinephrine or vasopressin.MethodsForty-one patients undergoing elective off-pump coronary artery bypass grafting participated in this study. After the median sternotomy, the left internal thoracic artery was dissected with a pedicle and grafted to the left anterior descending artery. After all anastomoses were performed and hemodynamic parameters were stable, the grafted internal thoracic arterial blood flow was measured by transit time flowmeter on the distal portion of the graft as a baseline. Norepinephrine or vasopressin was then infused until mean arterial pressure was increased to 20% of baseline. Graft flow and hemodynamic variables were measured when mean arterial pressure reached the intended level.ResultsBaseline grafted internal thoracic arterial flows were similar (norepinephrine 57.1 ± 17.7 mL min−1, vasopressin 66.0 ± 34.3 mL min−1). With norepinephrine, flow increased significantly relative to baseline (77.2 ± 31.0 mL min−1); with vasopressin, it remained unchanged (68.3 ± 37.0 mL min−1).ConclusionsFor patients needing vasopressor support after coronary artery bypass grafting, norepinephrine appeared superior to vasopressin because of increased internal thoracic arterial flow

Signal pathways in astrocytes activated by cross-talk between of astrocytes and mast cells through CD40-CD40L

<p>Abstract</p> <p>Background</p> <p>Astrocytes, which play an active role in chronic inflammatory diseases like multiple sclerosis, exist close to mast cells with which they share perivascular localization. We previously demonstrated the possibility that astrocytes and mast cells interact in vitro and in vivo. This study aimed to investigate the signaling pathways and the role for astrocytes in the interaction of astrocytes and mast cells.</p> <p>Methods</p> <p>We co-cultured human U87 glioblastoma (U87) and human mast cell-1 (HMC-1) cell lines, and mouse cerebral cortices-derived astrocytes and mouse bone marrow-derived mast cells (BMMCs). Intracellular Ca²⁺([Ca²⁺]_i) was measured by confocal microscopy; CD40 siRNA by Silencer Express Kit; small GTPases by GTP-pull down assay; PKCs, MAPKs, CD40, CD40L, Jak1/2, STAT1, TNF receptor 1 (TNFR1) by Western blot; NF-κB and AP-1 by EMSA; cytokines by RT-PCR. An experimental allergic encephalomyelitis (EAE) model was induced using myelin oligodendrocyte glycoprotein (MOG) peptide and pertussis toxin in mice. Co-localization of TNFR1 and astrocytes in EAE brain tissues was determined by immunohistochemistry.</p> <p>Results</p> <p>Each astrocyte co-culture had increases in [Ca²⁺]_ilevels, release of cytokines and chemokines; activities of Rho-family GTPases, NF-κB/AP-1/STAT1⁷²⁷, and Jack1/2, STAT1⁷⁰¹. These effects were inhibited by anti-CD40 antibody or CD40 siRNA, and signaling pathways for Jak1/2 were inhibited by anti-TNFR1 antibody. EAE score, expression of TNFR1, and co-localization of TNFR1 and astrocytes were enhanced in brain of the EAE model. Anti-CD40 antibody or 8-oxo-dG pretreatment reduced these effects in EAE model.</p> <p>Conclusions</p> <p>These data suggest that astrocytes activated by the CD40-CD40L interaction in co-culture induce inflammatory cytokine production via small GTPases, and the secreted cytokines re-activate astrocytes via Jak/STAT1⁷⁰¹pathways, and then release more cytokines that contribute to exacerbating the development of EAE. These findings imply that the pro-inflammatory mediators produced by cell-to-cell cross-talk via interaction of CD40-CD40L may be as a promising therapeutic target for neurodegenerative diseases like MS.</p