Nucleic acid analysis is one of the most important disease diagnostic approaches in medical practice, and has been commonly used in cancer biomarker detection, bacterial speciation and many other fields in laboratory. Currently, the application of powerful research methods for genetic analysis, including the polymerase chain reaction (PCR), DNA sequencing, and gene expression profiling using fluorescence microarrays, are not widely used in hospitals and extended-care units due to high-cost, long detection times, and extensive sample preparation. Bioassays, especially chip-based electrochemical sensors, may be suitable for the next generation of rapid, sensitive, and multiplexed detection tools. Herein, we report three different microelectrode platforms with capabilities enabled by nano- and microtechnology: nanoelectrode ensembles (NEEs), nanostructured microelectrodes (NMEs), and hierarchical nanostructured microelectrodes (HNMEs), all of which are able to directly detect unpurified RNA in clinical samples without enzymatic amplification. Biomarkers that are cancer and infectious disease relevant to clinical medicine were chosen to be the targets. Markers were successfully detected with clinically-relevant sensitivity. Using peptide nucleic acids (PNAs) as probes and an electrocatalytic reporter system, NEEs were able to detect prostate cancer-related gene fusions in tumor tissue samples with 100 ng of RNA. The development of NMEs improved the sensitivity of the assay further to 10 aM of DNA target, and multiplexed detection of RNA sequences of different prostate cancer-related gene fusion types was achieved on the chip-based NMEs platform. An HNMEs chip integrated with a bacterial lysis device was able to detect as few as 25 cfu bacteria in 30 minutes and monitor the detection in real time. Bacterial detection could also be performed in neat urine samples. The development of these versatile clinical diagnostic tools could be extended to the detection of various cancers, genetic, and infectious diseases.

Mitochondria play essential roles in numerous cellular processes, including oxidative phosphorylation and apoptotic initiation. As a result, organellar dysfunction has been implicated in several pathologies such as cancer, diabetes and neurodegenerative diseases. The opportunity to deliver compounds to probe or treat these conditions would be highly beneficial but accessing this organelle is challenging. Prior work investigated the physicochemical properties required for mitochondrial targeting and yielded mitochondria-penetrating peptides (MPPs). MPPs possess hydrophobic and cationic character and exhibit efficient cellular uptake and mitochondrial localization. In this proof-of-principle study, MPPs were harnessed to re-route an anti-leukemia agent, chlorambucil (Cbl), from the nucleus to mitochondria. This DNA alkylating agent was selected for its rapid kinetics and facile conjugation to an MPP. In addition, because mitochondria possess their own genome, the target of this drug would also be present in the organelle. Conjugation of an MPP to Cbl (mt-Cbl) confirmed that the drug was re-routed to the mitochondria and an increase in potency was observed in several cell lines and patient samples. This gain in activity was due to the increased accessibility of the mitochondrial genome, its lack of introns and its limited repair capacity. However, despite this enhanced toxicity, a therapeutic window continued to be maintained due to the elevated mitochondrial membrane potential in cancer cells. The re-routing of Cbl also resulted in evasion of several drug resistance mechanisms. Damage directly within the organelle was sufficient to initiate apoptosis even in cell lines with disabled apoptotic triggering. In addition, mitochondrial sequestration protected mt-Cbl from drug inactivation mechanisms. Lastly, mt-Cbl inhibited Pgp efflux by unexpectedly interacting with the pumps and inhibiting activity for a short period of time. The anti-cancer activity of mt-Cbl was also assessed in vivo in xenograft models of leukemia. The conjugate was stable in mouse plasma and displayed an improved pharmacokinetic profile. In addition, mt-Cbl successfully delayed tumor growth in two xenograft models and continued to alkylate mitochondrial DNA in vivo. These studies demonstrate that MPPs can be harnessed to re-route drugs to this organelle. Mitochondrial re-targeting could be a novel method of re-purposing FDA-approved drugs to enhance activity and evade resistance.

The use of multimodal imaging as a tool to assess the in vivo pharmacokinetics and biodistribution of nanoparticles is important in drug development and imaging-guided therapy. The current study reports the use of combined micro-CT and optical imaging FMT to quantitatively assess the whole body and intratumoural distribution of a nano-sized liposome-based CT/optical imaging probe carrying iohexol and Cy5.5. Micro-CT was used to guide the FMT tumour delineation and signal correction. This investigation demonstrates the critical role micro-CT can play in guiding FMT-based quantification of distribution. As well the combination of CT and optical imaging enable visualization of the liposomes at the whole body, tumor and cellular levels with high sensitivity and excellent anatomical background. Such non-invasive assessment of therapeutic distribution at the macro and micro scale is necessary for implementation of personalized medicine including image-guided patient stratification and real-time adjustment of therapeutic dose.

Objective: To reach consensus on entry-to-practice natural health product (NHP)-related competency statements for Canadian pharmacy students. Methods: Four rounds of a modified Delphi method were conducted. Participants; pharmacy educators,and representatives from Canadian pharmacy organizations (n=17), ranked their level of agreement using a 5-point Likert scale. Results: Consensus occurred when all participants ranked a statement 4 or 5. Three core NHP-related competencies were identified: 1) the ability to incorporate NHP knowledge when providing pharmaceutical care; 2) the ability to access and critically appraise NHP-related information sources, and 3) the ability to provide appropriate education to patients and other health care providers on the effectiveness and potential adverse effects and drug interactions of NHPs. Two additional NHP-related competency statements emerged as important, but consensus was not achieved. Conclusions: If the developed core NHP-related competency statements are widely implemented, Canadian pharmacists will be able to fulfill their NHP-related professional responsibilities upon entry-to-practice.

Background: Although many pharmacies sell natural health products (NHPs), there is no clear definition as to the responsibilities (if any) of pharmacists towards these products. Objective: The purpose of this study was to explore and compare pharmacy and stakeholder leaders’ perceptions of pharmacists’ professional NHP responsibilities. Methods: Semi-structured key informant interviews were conducted with pharmacy leaders and stakeholder leaders representing: consumers, complementary and alternative medicine practitioners, conventional healthcare practitioners, and industry across Canada. Results: Nearly all participants believed safety monitoring was a key responsibility of pharmacists. One challenge identified was pharmacists’ general lack of NHP knowledge. Stakeholder leaders did not expect pharmacists to be NHP experts, but should have a basic level of education on NHPs. Many pharmacy leaders seemed unfamiliar with current pharmacy NHP policies. Conclusion: Participants described pharmacists’ professional responsibilities for NHPs as similar to those for over-the-counter drugs. More awareness of existing NHP-related pharmacy policies is needed.

Targeting angiogenic vasculature has been validated as a viable approach for cancer imaging and therapy. The tumour vasculature-specific ligand asparagine-glycine-arginine (NGR) peptide targets the isoform of aminopeptidase N (CD13) expressed on endothelial cells lining angiogenic vessels. CD13 has become widely recognized as a rational target for therapeutic development and several NGR-conjugated agents are now in pre-clinical and clinical development. In the current study, a CT image-based approach is used to evaluate the in vivo performance of several NGR-conjugated liposome formulations that vary in terms of NGR density and PEG spacer arm length. Indeed, for the first time it is demonstrated that CT imaging can be used for quantitative and longitudinal assessment of the pharmacokinetics and biodistribution of an actively targeted liposome formulation. In comparison to conventional methods, CT imaging enables visualization of the intratumoural distribution of liposomes and quantification of the fraction of tumour occupied by the vesicles over time.

Objectives: To use the means-end chain (MEC) decision-making approach to compare two groups of participants using natural health products (NHPs) with and without scientific evidence support. Methods: The laddering technique was used to interview 25 participants with osteoarthritis. Hierarchical value maps were generated to depict the decision-making processes. Semi-structured questions probed the role of scientific evidence in the decision-making process and content analysis identified thematic similarities and differences between the two groups. Results: The dominant decision-making chain between participants in the two scientific evidence categories was similar. Scientific evidence is an important decision-making factor but not as important as the advice from health care providers, friends and family. Conclusions: The MEC-approach and its associated laddering methodology helped us understand how people make decisions about NHPs. There were essentially no differences in how consumers in our two groups incorporated scientific evidence into their choice of NHPs for OA.

Targeting angiogenic vasculature has been validated as a viable approach for cancer imaging and therapy. The tumour vasculature-specific ligand asparagine-glycine-arginine (NGR) peptide targets the isoform of aminopeptidase N (CD13) expressed on endothelial cells lining angiogenic vessels. CD13 has become widely recognized as a rational target for therapeutic development and several NGR-conjugated agents are now in pre-clinical and clinical development. In the current study, a CT image-based approach is used to evaluate the in vivo performance of several NGR-conjugated liposome formulations that vary in terms of NGR density and PEG spacer arm length. Indeed, for the first time it is demonstrated that CT imaging can be used for quantitative and longitudinal assessment of the pharmacokinetics and biodistribution of an actively targeted liposome formulation. In comparison to conventional methods, CT imaging enables visualization of the intratumoural distribution of liposomes and quantification of the fraction of tumour occupied by the vesicles over time.

Objectives: To use the means-end chain (MEC) decision-making approach to compare two groups of participants using natural health products (NHPs) with and without scientific evidence support. Methods: The laddering technique was used to interview 25 participants with osteoarthritis. Hierarchical value maps were generated to depict the decision-making processes. Semi-structured questions probed the role of scientific evidence in the decision-making process and content analysis identified thematic similarities and differences between the two groups. Results: The dominant decision-making chain between participants in the two scientific evidence categories was similar. Scientific evidence is an important decision-making factor but not as important as the advice from health care providers, friends and family. Conclusions: The MEC-approach and its associated laddering methodology helped us understand how people make decisions about NHPs. There were essentially no differences in how consumers in our two groups incorporated scientific evidence into their choice of NHPs for OA.

Background: Although many pharmacies sell natural health products (NHPs), there is no clear definition as to the responsibilities (if any) of pharmacists towards these products. Objective: The purpose of this study was to explore and compare pharmacy and stakeholder leaders’ perceptions of pharmacists’ professional NHP responsibilities. Methods: Semi-structured key informant interviews were conducted with pharmacy leaders and stakeholder leaders representing: consumers, complementary and alternative medicine practitioners, conventional healthcare practitioners, and industry across Canada. Results: Nearly all participants believed safety monitoring was a key responsibility of pharmacists. One challenge identified was pharmacists’ general lack of NHP knowledge. Stakeholder leaders did not expect pharmacists to be NHP experts, but should have a basic level of education on NHPs. Many pharmacy leaders seemed unfamiliar with current pharmacy NHP policies. Conclusion: Participants described pharmacists’ professional responsibilities for NHPs as similar to those for over-the-counter drugs. More awareness of existing NHP-related pharmacy policies is needed.