The synthesis and validation of multimodal PET/fluorescence zinc sensing probes as potential imaging agents for prostate cancer

Prostate cancer (PCa) is the 2nd most common cancer worldwide for males, and the 4th most common cancer overall, accounting for 15% of all male cancers in 2012. PCa is asymptomatic in its early stages, advances in medical imaging are improving diagnosis, however novel simple medical imaging agents are needed to diagnose the disease early. A fluorescent imaging probe, 2-((2-(2-fluoroethoxy)ethyl)amino)-N-(quinolin-8-yl)acetamide (AQA-F), that changes emission profile when binding the metal zinc has been synthesised. A Stokes shift of 80 nm can be observed upon zinc binding, enhancing the emission wavelength from 420 to 500 nm, with a Kd of 14x10⁻⁶ M. It was shown that AQA-F has the highest affinity for Zn²⁺ which will allow for the in vitro differentiation of zinc bound and free probe. AQA-F was non-toxic at imaging concentrations in healthy prostate, prostate cancer and healthy tissues. It was also shown that AQA-F was internalised by healthy and prostate cancer cell lines across both 2D and 3D models. This allows for the probe to function as an endogenous zinc sensor in vitro with the potential of being translated into clinical use for the diagnosis of prostate cancer in men. Further studies will be required to identify a radiolabelling procedure for the probe with ¹⁸F to enable possible dual modal imaging

Offshore Outsourcing and Organizational Learning: A Model of Cultural Occlusions

Offshore outsourcing of information technology services is a recent global trend. Organizations may not have considered the impact of outsourcing on their learning capabilities for future competitive competencies. The impact on learning may depend on the national cultures of the client organization and of the offshore outsourcing vendor. A conceptual model of cultural occlusions to organizational learning in IT outsourcing relationships is developed in this paper. Suggested solutions include employing boundary spanners to mitigate of any unfavourable impacts. The cultural occlusions model may inform IT practitioners in an offshore outsourcing relationship, and the IT industry strategy of developing countries offering offshore outsourcing services

Bacterial outer-membrane vesicles promote Vγ9Vδ2 T cell oncolytic activity

BACKGROUND: Increasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as γδ T cells has been suggested. METHODS: Peripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Δpal ΔlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated γδ T cells were co-cultured with cancer cell lines to determine cytotoxicity. RESULTS: The vesicles induced a broad inflammatory response with γδ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of γδ T cells were of the Vγ9Vδ2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of Vγ9Vδ2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion. CONCLUSIONS: Our findings identify for the first time, that OMV-challenge stimulates the expansion of Vγ9Vδ2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of Vγ9Vδ2 T cells, an axis amenable for improved future therapeutics

Drug-mediated shortening of action potentials in LQTS2 hiPSC-cardiomyocytes

Cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSCs) are now a well-established modality for modeling genetic disorders of the heart. This is especially so for long QT syndrome (LQTS), which is caused by perturbation of ion channel function, and can lead to fainting, malignant arrhythmias and sudden cardiac death. LQTS2 is caused by mutations in KCNH2, a gene whose protein product contributes to IKr (also known as HERG), which is the predominant repolarizing potassium current in CMs. β-blockers are the mainstay treatment for patients with LQTS, functioning by reducing heart rate and arrhythmogenesis. However, they are not effective in around a quarter of LQTS2 patients, in part, because they do not correct the defining feature of the condition, which is excessively prolonged QT interval. Since new therapeutics are needed, in this report, we biopsied skin fibroblasts from a patient who was both genetically and clinically diagnosed with LQTS2. By producing LQTS-hiPSC-CMs, we assessed the impact of different drugs on action potential duration (APD), which is used as an in vitro surrogate for QT interval. Not surprisingly, the patient's own β-blocker medication, propranolol, had a marginal effect on APD in the LQTS-hiPSC-CMs. However, APD could be significantly reduced by up to 19% with compounds that enhanced the IKr current by direct channel binding or by indirect mediation through the PPARδ/protein 14-3-3 epsilon/HERG pathway. Drug-induced enhancement of an alternative potassium current, IKATP, also reduced APD by up to 21%. This study demonstrates the utility of LQTS-hiPSC-CMs in evaluating whether drugs can shorten APD and, importantly, shows that PPARδ agonists may form a new class of therapeutics for this condition

A 18F radiolabelled Zn(ii) sensing fluorescent probe

A selective fluorescent probe for Zn(ii), AQA-F, has been synthesized. AQA-F exhibits a ratiometric shift in emission of up to 80 nm upon binding Zn(ii) ([AQA-F] = 0.1 mM, [Zn(ii)Cl 2 ] = 0-300 μM). An enhancement of quantum yield from Φ = 4.2% to Φ = 35% is also observed. AQA-F has a binding constant, K d = 15.2 μM with Zn(ii). This probe has been shown to respond to endogenous Zn(ii) levels in vitro in prostate and prostate cancer cell lines. [ 18 F]AQA-F has been synthesized with a radiochemical yield of 8.6% and a radiochemical purity of 97% in 88 minutes. AQA-F shows the potential for a dual modal PET/fluorescence imaging probe for Zn(ii)

Design and fabrication of instrumented composite airfoils for a cryogenic wind tunnel model

Two instrumented horizontal stabilizers and one instrumented vertical stabilizer were designed and fabricated for testing on the Pathfinder 1 (PF-1) Transport Model in the NASA Langley Research Center's National Transonic Facility (NTF). Two different designs were employed: the horizontal stabilizer utilized a metal spar and fiberglass overwrap and the vertical stabilizer was made of all fiberglass. All design requirements were met in terms of design loads, airfoil tolerances, surface finish, orifice hole quality, and proof-of-concept tests. Pressure tubing installation was found to be easier for these concepts as compared to methods used in conventional metallic models. Ease of repair was found to be a principal advantage in that some fabrication problems were overcome by reapplying fiberglass cloth and/or epoxy to damaged areas. Also, fabrication costs were judged to be lower when compared to the more conventional design fabrication costs

Receptor-targeted peptide conjugates based on diphosphines enable preparation of 99mTc and 188Re theranostic agents for prostate cancer

Benchtop 99Mo/ 99mTc and 188W/ 188Re generators enable economical production of molecular theranostic 99mTc and 188Re radiopharmaceuticals, provided that simple, kit-based chemistry exists to radiolabel targeting vectors with these radionuclides. We have previously described a diphosphine platform that efficiently incorporates 99mTc into receptor-targeted peptides. Here, we report its application to label a prostate-specific membrane antigen (PSMA)-targeted peptide with 99mTc and 188Re for diagnostic imaging and systemic radiotherapy of prostate cancer. Methods: Two diphosphine-dipeptide bioconjugates, DP1-PSMAt and DP2-PSMAt, were formulated into kits for radiolabeling with 99mTc and 188Re. The resulting radiotracers were studied in vitro, in prostate cancer cells, and in vivo in mouse xenograft models, to assess similarity of uptake and biodistribution for each 99mTc/ 188Re pair of agents. Results: Both DP1-PSMAt and DP2-PSMAt could be efficiently radiolabeled with 99mTc and 188Re using kit-based methods to furnish the isostructural compounds M-DP1-PSMAt and M-DP2-PSMAt (M = [ 99mTc]Tc, [ 188Re]Re). All 99mTc/ 188Re radiotracers demonstrated specific uptake in PSMA-expressing prostate cancer cells, with negligible uptake in prostate cancer cells that did not express PSMA or in which PSMA uptake was blocked. M-DP1-PSMAt and M-DP2-PSMAt also exhibited high tumor uptake (18-30 percentage injected dose per gram at 2 h after injection), low retention in nontarget organs, fast blood clearance, and excretion predominantly via a renal pathway. Importantly, each pair of 99mTc/ 188Re radiotracers showed near-identical biologic behavior in these experiments. Conclusion: We have prepared and developed novel pairs of isostructural PSMA-targeting 99mTc/ 188Re theranostic agents. These generator-based theranostic agents have potential to provide access to the benefits of PSMA-targeted diagnostic imaging and systemic radiotherapy in health care settings that do not routinely have access to either reactor-produced 177Lu radiopharmaceuticals or PET/CT infrastructure. </p

Platelet zinc status regulates prostaglandin-induced signaling, altering thrombus formation

Background: Approximately 17.3% of the global population exhibits an element of zinc (Zn2+) deficiency. One symptom of Zn2+ deficiency is increased bleeding through impaired hemostasis. Platelets are crucial to hemostasis and are inhibited by endothelial-derived prostacyclin (prostaglandin I2 [PGI2]), which signals via adenylyl cyclase (AC) and cyclic adenosine monophosphate signaling. In other cell types, Zn2+ modulates cyclic adenosine monophosphate concentrations by changing AC and/or phosphodiesterase activity. Objectives: To investigate if Zn2+ can modulate platelet PGI2 signaling. Methods: Platelet aggregation, spreading, and western blotting assays with Zn2+ chelators and cyclic nucleotide elevating agents were performed in washed platelets and platelet-rich plasma conditions. In vitro thrombus formation with various Zn2+ chelators and PGI2 was assessed in whole blood. Results: Incubation of whole blood or washed platelets with Zn2+ chelators caused either embolization of preformed thrombi or reversal of platelet spreading, respectively. To understand this effect, we analyzed resting platelets and identified that incubation with Zn2+ chelators elevated pVASPser157, a marker of PGI2 signaling. In agreement that Zn2+ affects PGI2 signaling, addition of the AC inhibitor SQ22536 blocked Zn2+ chelation–induced platelet spreading reversal, while addition of Zn2+ blocked PGI2-mediated platelet reversal. Moreover, Zn2+ specifically blocked forskolin-mediated AC reversal of platelet spreading. Finally, PGI2 inhibition of platelet aggregation and in vitro thrombus formation was potentiated in the presence of low doses of Zn2+ chelators, increasing its effectiveness in inducing platelet inhibition. Conclusion: Zn2+ chelation potentiates platelet PGI2 signaling, elevating PGI2’s ability to prevent effective platelet activation, aggregation, and thrombus formation