P2Y1 receptor modulation of endogenous ion channel function in Xenopus oocytes: Involvement of transmembrane domains

Agonist activation of the hP2Y1 receptor expressed in Xenopus oocytes stimulated an endogenous voltage-gated ion channel, previously identified as the transient inward (Tin) channel. When human P2Y1 (hP2Y1) and skate P2Y (sP2Y) receptors were expressed in Xenopus oocytes, time-to-peak values (a measure of the response to membrane hyperpolarization) of the Tin channel were significantly reduced compared to oocytes expressing the hB1-bradykinin receptor or the rat M1-muscarinic (rM1) receptor. Differences in activation were also observed in the Tin currents elicited by various P2Y receptor subtypes. The time-to-peak values of the Tin channel in oocytes expressing the hP2Y4, hP2Y11, or hB1-bradykinin receptors were similar, whereas the channel had significantly shorter time-to-peak values in oocytes expressing either the hP2Y1 or sP2Y receptor. Amino acid substitutions at His-132, located in the third transmembrane domain (TM3) of the hP2Y1 receptor, delayed the onset of channel opening, but not the kinetics of the activation process. In addition, Zn2+ sensitivity was also dependent on the subtype of P2Y receptor expressed. Replacement of His-132 in the hP2Y1 receptor with either Ala or Phe increased Zn2+ sensitivity of the Tin current. In contrast, truncation of the C-terminal region of the hP2Y1 receptor had no affect on activation or Zn2+ sensitivity of the Tin channel. These results suggested that TM3 in the hP2Y1 receptor was involved in modulating ion channel function and blocker pharmacology of the Tin channel

Analysis of shared heritability in common disorders of the brain

Paroxysmal Cerebral Disorder

Considering planned change anew: stretching large group interventions strategically, emotionally and meaningfully

Large Group Interventions, methods for involving “the whole system” in a change process, are important contemporary planned organizational change approaches. They are well known to practitioners but unfamiliar to many organizational researchers, despite the fact that these interventions address crucial issues about which many organizational researchers are concerned. On the other hand, these interventions do not appear to be informed by contemporary developments in organizational theorizing. This disconnect on both sides is problematic. We describe such interventions and their importance; illustrate them with extended descriptions of particular Future Search and Whole‐Scale™ change interventions; summarize research on strategy, emotion, and sensemaking that may inform them; and suggest questions about the interventions that may stimulate research and reflection on practice. We also discuss conditions that may foster effective engagement between Large Group Interventions practitioners and organizational researchers. Our approach represents a way to conduct a review that combines scholarly literature and skilled practice and to initiate a dialog between them

Detection of Chlamydia trachomatis in genital swabs: comparison of commercial and in house amplification methods with culture

AIMS: To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies (EB) and also to test 245 urethral and endocervical specimens for Chlamydia trachomatis by the four assays as well as conventional culture. STUDY DESIGN: EB titrations were run in duplicate in each commercial assay and six times in the in house PCR. Clinical samples were aliquoted and tested by each assay and were considered positive if C trachomatis was detected by two or more separate tests or if the sample was either culture or immunofluorescence positive. Major outer membrane protein (MOMP) specific primers were used as a confirmatory assay for the in house PCR. RESULTS: The in house PCR, Roche Cobas Amplicor, LCR, and Amplicor plate kit gave detection limits of approximately 1, 1-2, 2, and 2-4 EBs respectively. By the criteria described above for definition of a C trachomatis positive result in clinical samples we identified 23 true positives among the 245 clinical specimens. The in house PCR detected all 23 giving a sensitivity of 100% and a specificity of 98%. The Roche Cobas Amplicor, Roche Amplicor plate kit, and LCR detected 21, 19, and 19 of these respectively giving sensitivities of 87.5%, 82%, and 82% respectively and specificities of 99.5%, 99%, and 100% respectively. The culture gave a sensitivity of 78% and specificity of 100%. CONCLUSION: All four amplification assays had a greater sensitivity than the culture used routinely in this laboratory. The in house plasmid PCR had the greatest sensitivity and when combined with confirmation by immunofluorescence detected the greatest number of positives. This increased sensitivity is likely to have been achieved by the use of a DNA purification step and of nested primers in the amplification stage and their combined use in routine diagnostic assays for chlamydia might increase the frequency of C trachomatis detections. However, this assay is much less user friendly than the two semiautomated commercial assays investigated in this study. 



TIL therapy broadens the tumor-reactive CD8(+) T cell compartment in melanoma patients

Toxicolog

Phase III randomized clinical trial comparing tremelimumab with standard-of-care chemotherapy in patients with advanced melanoma

Purpose In phase I/II trials, the cytotoxic T lymphocyte-associated antigen-4-blocking monoclonal antibody tremelimumab induced durable responses in a subset of patients with advanced melanoma. This phase III study evaluated overall survival (OS) and other safety and efficacy end points in patients with advanced melanoma treated with tremelimumab or standard-of-care chemotherapy. Patients and Methods Patients with treatment-naive, unresectable stage IIIc or IV melanoma were randomly assigned at a ratio of one to one to tremelimumab (15 mg/kg once every 90 days) or physician's choice of standard-of-care chemotherapy (temozolomide or dacarbazine). Results In all, 655 patients were enrolled and randomly assigned. The test statistic crossed the prespecified futility boundary at second interim analysis after 340 deaths, but survival follow-up continued. At final analysis with 534 events, median OS by intent to treat was 12.6 months (95% CI, 10.8 to 14.3) for tremelimumab and 10.7 months (95% CI, 9.36 to 11.96) for chemotherapy (hazard ratio, 0.88; P = .127). Objective response rates were similar in the two arms: 10.7% in the tremelimumab arm and 9.8% in the chemotherapy arm. However, response duration (measured from date of random assignment) was significantly longer after tremelimumab (35.8 v 13.7 months; P = .0011). Diarrhea, pruritus, and rash were the most common treatment-related adverse events in the tremelimumab arm; 7.4% had endocrine toxicities. Seven deaths in the tremelimumab arm and one in the chemotherapy arm were considered treatment related by either investigators or sponsor. Conclusion This study failed to demonstrate a statistically significant survival advantage of treatment with tremelimumab over standard-of-care chemotherapy in first-line treatment of patients with metastatic melanoma. © 2013 by American Society of Clinical Oncology

Building a Sibling Aggression Treatment Model: Design and Development Research in Action

One strategy for addressing the persistent challenge of advancing empirically based social work practice is to engage practitioners in the formulation of empirically developed practice guidelines. Although this approach is promising, there has been relatively little work to guide practitioners, and perhaps consequently, few examples of practitioner-developed guidelines exist. Moreover, available treatment models do not address most social work practice concerns. One such concern involves sibling aggression, which is exceptionally widespread with known deleterious consequences, but it has received scant attention in the intervention literature. This article illustrates a practitioner\u27s development of a task-centered sibling aggression treatment model using the design and development research paradigm, a methodology for systematically formulating, testing, and refining practice guidelines. It also introduces a promising sibling aggression treatment model