Presenilin-1 is required for γ-secretase activity, which participates in Notch receptor processing, the pathogenesis of Alzheimer's disease and the modulation of Ca2+ signaling. We tested the hypothesis that γ-secretase proteolytic activity modulates store-operated Ca2+ entry (SOCE) in rat dorsal root ganglion neurons. Depletion of intracellular Ca2+ stores by blocking the endoplasmic reticulum Ca2+ pump with cyclopiazonic acid (CPA) evoked a transient increase in [Ca2+]i but no sustained Ca2+ influx. However, in cells expressing a dominant negative presenilin-1 mutant (PS1-D257A), γ-secretase activity was inhibited and treatment with CPA evoked sustained Ca2+ influx. Similarly, pharmacologic inhibition of γ-secretase with DAPT for 48 hrs enhanced SOCE. SKF96365, an inhibitor of store-operated channels, blocked SOCE in cells expressing PS1-D257A. Thus, γ-secretase proteolytic activity regulates a SOCE pathway in sensory neurons.

Microglia, the resident macrophages of the CNS, are responsible for the innate immune response in the brain and participate in the pathogenesis of certain neurodegenerative disorders. Chemokines initiate activation and migration of microglia. The beta-chemokine CCL5 induces an elevation in intracellular calcium concentration ([Ca(2+)](i)) in human microglia. Here, we examined the signal transduction pathway linking activation of chemokine receptor CCR5 to an elevation in [Ca(2+)](i) in cultured microglia by using pharmacological approaches in combination with Fura-2-based digital imaging. The CCL5-induced response required Janus kinase (Jak) activity and the stimulation of an inhibitory G protein. Multiple downstream signaling pathways were involved, including phosphatidylinositol 3-kinase (PI3K), Bruton's tyrosine kinase (Btk), and phospholipase C (PLC)-mediated release of Ca(2+) from inositol 1,4,5-trisphosphate (IP(3))-sensitive stores. Activation of both the kinase and the lipase pathways was required for eliciting the Ca(2+) response. However, the majority of the [Ca(2+)](i) increase was derived from sources activated by NAD metabolites. Cyclic ADP-ribose (cADPR) evoked Ca(2+) release from intracellular stores, and ADPR evoked Ca(2+) influx via a nimodipine-sensitive channel. Thus, a multistep cascade couples CCR5 activation to Ca(2+) increases in human microglia. Because changes in [Ca(2+)](i) affect chemotaxis, secretion, and gene expression, pharmacologic modulation of this pathway may alter inflammatory and degenerative processes in the CNS.

Optimal management for unplanned excision of soft-tissue sarcoma is unknown. Our institution has adopted the approach of preoperative radiotherapy, followed by definitive surgery. In our series of 44 patients, local control was excellent at 95%, with perioperative complications seen only in patients with lower extremity sarcomas, suggesting that this is a reasonable approach to manage inadvertently resected sarcoma.

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institution, along with early results of the treated patients. It is an attempt to present potential challenges and solutions applicable for radiation oncology departments in developed and developing nations alike. Materials/Methods: We describe the 4-month multidisciplinary planning phase to initiate an IGBT program for cervical cancer. We describe the specific challenges that were encountered prior to treating our first patient, and we also report on the outcome of the treated patients between November 2015 and December 2017. Results: We describe the solutions that were realized and executed to solve the clinical and physics challenges that we faced to establish IGBT program within the context of our available institutional resources. In the planning phase we established a team with defined job description for each member. We emphasize detailed coordination of care, planning, and communication to make the workflow feasible, and ensure proper education of the team members and collaborators from other departments about every aspect of IGBT. We present the imaging and radiation physics solutions to ensure a safe delivery of IGBT before having the first patient on table. For this purpose, a 3D brachytherapy plan with just the applicator was created, delivered and analyzed using radiographic films. CT based planning was used for all the fractions. Due to limited resources, one MRI was obtained for target volume definition with the applicator in situ for the first fraction, and fused with subsequent CTs for the remaining fractions. Volume definition and brachytherapy planning were in accordance with GEC-ESTRO guidelines for 3D IGBT. The strategies that were developed to overcome the challenges and the experience that was built over the time represented a learning curve for all the members, thus resulting in a decrease of the total time from about 7 hours to 4 hours for each fraction. Nine patients were treated so far with a median follow up time of 11 months (range 6 e 17). The median external beam radiotherapy dose was 50.4Gy with concurrent Cisplatin, and the brachytherapy dose was 28Gy/ 4Fx. 7 patients achieved complete remission in the primary, but one of them died of liver metastases 7 months after treatment completion. Of the remaining two, one died of progressive local and distant disease soon after treatment completion, and one is alive with evidence of disease in the pelvis. None of these 9 patients developed more than grade 2 GI or GU toxicities Conclusion: This review represents a detailed report on establishing a safe and efficient 3D IGBT service, and it addresses important lessons learned from a successful establishment of a sophisticated radio-therapeutic modality in developing countries.