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The use of benign stimuli to control the binding and release of labile biologics for their isolation from complex feedstocks is a key goal of modern biopharmaceutical technology. This study introduces cyclic azobenzene-peptide (CAP) hybrid ligands for the rapid and discrete photo-responsive capture and release of blood coagulation Factor VIII (FVIII). A predictive method - based on amino acid sequence and molecular architecture of CAPs - was developed to correlate the conformation of cis/trans CAP photo-isomers to FVIII binding and release. The combined in silico and in vitro analysis of FVIII:peptide interactions guided the design of a rational approach to optimize isomerization kinetics and biorecognition of CAPs. A photoaffinity adsorbent, prepared by conjugating selected CAP G-cyclo AZOB [Lys-YYKHLYN-Lys]-G on translucent chromatographic beads, featured high binding capacity (> 6 mg of FVIII per mL of resin) and rapid photo-isomerization kinetics ( τ < 30s) when exposed to 420-450 nm light at the intensity of 0.1 W·cm -2 . The adsorbent purified FVIII from a recombinant harvest using a single mobile phase, affording high product yield (>90%), purity (>95%), and blood clotting activity. The CAPs introduced in this report demonstrate a novel route integrating gentle operational conditions in a rapid and efficient bioprocess for the purification of life-saving biotherapeutics., Competing Interests: Conflict of interest disclosure. No conflict of interest to declare.

Bilaterally symmetric flowers have evolved over a hundred times in angiosperms, yet orthologs of the transcription factors CYCLOIDEA ( CYC ), RADIALIS ( RAD ), and DIVARICATA ( DIV ) are repeatedly implicated in floral symmetry changes. We examined these candidate genes to elucidate the genetic underpinnings of floral symmetry changes in florally diverse Rhododendron , reconstructing gene trees and comparing gene expression across floral organs in representative species with radial and bilateral flower symmetries. Radially symmetric R. taxifolium Merr. and bilaterally symmetric R. beyerinckianum Koord. had four and five CYC orthologs, respectively, from shared tandem duplications. CYC orthologs were expressed in the longer dorsal petals and stamens and highly expressed in R. beyerinckianum pistils, whereas they were either ubiquitously expressed, lost from the genome, or weakly expressed in R. taxifolium . Both species had two RAD and DIV orthologs uniformly expressed across all floral organs. Differences in gene structure and expression of Rhododendron RAD compared to other asterids suggest that these genes may not be regulated by CYC orthologs. Our evidence supports CYC orthologs as the primary regulators of differential organ growth in Rhododendron flowers, while also suggesting certain deviations from the typical asterid gene regulatory network for flower symmetry.

Native platelets are crucial players in wound healing. Key to their role is the ability of their surface receptor GPIIb/IIIa to bind fibrin at injury sites, thereby promoting clotting. When platelet activity is impaired as a result of traumatic injury or certain diseases, uncontrolled bleeding can result. To aid clotting and tissue repair in cases of poor platelet activity, our lab has previously developed synthetic platelet-like particles capable of promoting clotting and improving wound healing responses. These are constructed by functionalizing highly deformable hydrogel microparticles (microgels) with fibrin-binding ligands including a fibrin-specific whole antibody or a single-domain variable fragment. To improve the translational potential of these clotting materials, we explored the use of fibrin-binding peptides as cost-effective, robust, high-specificity alternatives to antibodies. Herein, we present the development and characterization of soft microgels decorated with the peptide AHRPYAAK that mimics fibrin knob 'B' and targets fibrin hole 'b'. These "Fibrin-Affine Microgels with Clotting Yield" (FAMCY) were found to significantly increase clot density in vitro and decrease bleeding in a rodent trauma model in vivo . These results indicate that FAMCYs are capable of recapitulating the platelet-mimetic properties of previous designs while utilizing a less costly, more translational design., Competing Interests: Conflict of Interest Statement: Drs. Brown and Barker are Founders of SelSym Biotech, Inc. which is focused on developing injectable hemostatic materials.

Disseminated intravascular coagulation (DIC) is a pathological coagulopathy associated with infection that increases mortality. In DIC, excessive thrombin generation causes symptoms from formation of microthrombi to multiorgan failure; bleeding risks can also be a concern because of clotting factor consumption. Different clinical events lead to DIC, including sepsis, trauma, and shock. Treatments for thrombotic episodes or bleeding presentation in DIC oppose each other, thus creating therapeutic dilemmas in management. The objective of this study was to develop fibrin-specific core-shell nanogels (FSNs) loaded with tissue-type plasminogen activator (tPA) to treat the microcirculatory complications of DIC, which would facilitate targeted clot dissolution to manage microthrombi and the potential consumptive coagulopathy that causes bleeding. FSNs enhance formation of actively polymerizing clots by crosslinking fibrin fibers, but they can also target preexisting microthrombi and, when loaded with tPA, facilitate targeted delivery to lyse the microthrombi. We hypothesized that this dual action would simultaneously address bleeding and microthrombi with DIC to improve outcomes. In vivo, tPA-FSNs decreased the presentation of multiorgan microthrombi, recovered platelet counts, and improved bleeding outcomes in a DIC rodent model. When incorporated with human DIC patient plasma, tPA-FSNs restored clot structure and clot growth under flow. Together, these data demonstrate that a fibrinolytic agent loaded into fibrin-targeting nanogels could improve DIC outcomes., (© 2021 by The American Society of Hematology.)

Background: To optimize patient education, it is important to understand what healthcare professionals perceive to be ideal oncology medication education for patients to receive, and what they feel is their role and the role of others in its delivery. Education provided to patients is an important component of chemotherapy as it has been shown to benefit and positively impact patients who receive it. Educational interventions are often provided by multidisciplinary teams with the goal of improving patient care. However, few studies have explored the roles of healthcare professionals in delivering oncology medication education., Objective: To explore the perspectives of healthcare professionals working in medical, gynaecological or hematological oncology to identify what they perceive to be optimal oncology medication education for patients., Methods: Healthcare professionals (physicians, nurses and pharmacists) working in medical, gynaecological or hematological oncology at the Nova Scotia Health Authority, Central Zone were invited to participate in one-on-one, semi-structured interviews which were audio-recorded, transcribed and analyzed using thematic analysis., Findings: Fifteen interviews, including five physicians, four nurses and six pharmacists were conducted from February to April 2018. Four major themes were identified: Delivery of oncology medication education, Facilitating the patient learning process, Multidisciplinary Approach and Understanding barriers to the healthcare professional in providing education., Conclusion: The identified themes uncovered novel ideas about how healthcare professionals felt oncology medication education could ideally be delivered to patients, and supported findings in the literature. Although participants discussed barriers to their ability to deliver optimal education, they also identified ways in which they can facilitate patient learning, for example, through the reinforcement of education. Participants recognized the importance of increasing collaboration and communication with the multidisciplinary team. This research will inform the design of any new models for oncology medication education at the Nova Scotia Health Authority, Central Zone and potentially other sites., Competing Interests: The authors have declared that no competing interests exist.

Background: The scope of pharmacist practice has expanded in many jurisdictions, including Nova Scotia, Canada, to include prescribing of medications., Objective: To identify the relationship between barriers and facilitators to pharmacist prescribing and self-reported prescribing activity using the Theoretical Domains Framework version 2 (TDF(v2))., Methods: The study was a self-administered electronic survey of all registered pharmacists (approximately 1300) in Nova Scotia, Canada. The questionnaire was developed using a consensus process that mapped facilitators and barriers to prescribing with the 14 domains of the TDF(v2). The questionnaire captured information about the type and rate of pharmacists' prescribing activities, pharmacists' perceptions of their prescribing role at the patient, pharmacist, pharmacy organization and health system level, and pharmacist demographics and practice settings. A 5-point Likert scale was used for most TDF(v2) domains. Cronbach's alpha was used to study the internal consistency of responses within each of the TDF(v2) domains and simple logistic regression was used to measure the relationship between TDF(v2) domain responses and self-reported prescribing activity. Open-ended questions were analyzed separately., Results: Eighty-seven pharmacists completed the questionnaire. The majority of respondents were female (70 %), staff pharmacists (52 %) practicing pharmacy for a mean of 18 years. The three domains that respondents most positively associated with prescribing were Knowledge (84 %), Reinforcement (81 %) and Intentions (78 %). The largest effect on prescribing activity was the Skills domain (OR 4.41, 95% CI, 1.34-14.47)., Conclusions: We determined the TDF(v2) domains associated with pharmacist self-reported prescribing behaviours. This understanding can assist the development of policy and program interventions at the pharmacist, pharmacy, and health system levels, to increase the uptake of pharmacist prescribing. Further work is needed to develop and implement interventions based on the domains identified, and to test these in pilot settings and then in large-scale interventions., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: In hospitals around the world, there has been no consensus regarding which clinical activities a pharmacist should focus on until recently. In 2011, a Canadian clinical pharmacy key performance indicator (cpKPI) collaborative was formed. The goal of the collaborative was to advance pharmacy practice in order to improve patient outcomes and enhance the quality of care provided to patients by hospital pharmacists. Following a literature review, which indicated that pharmacists can improve patient outcomes by carrying out specific activities, and an evidence-informed consensus process, a final set of eight cpKPIs were established. Canadian hospitals leading the cpKPI initiative are currently in the early stages of implementing these indicators., Objective: To explore pharmacists' perceptions of the barriers and facilitators to the implementation of cpKPIs., Methods: Clinical pharmacists employed by the Nova Scotia Health Authority were invited to participate in focus groups. Focus group discussions were audio-recorded and transcribed, and data was analyzed using thematic analysis., Findings: Three focus groups, including 26 pharmacists, were conducted in February 2015. Three major themes were identified. Resisting the change was comprised of documentation challenges, increased workload, practice environment constraints, and competing priorities. Embracing cpKPIs was composed of seeing the benefit, demonstrating value, and existing supports. Navigating the unknown was made up of quality versus quantity battle, and insights into the future., Conclusions: Although pharmacists were challenged by documentation and other changes associated with the implementation of cpKPIs, they demonstrated significant support for cpKPIs and were able to see benefits of the implementation. Pharmacists came up with suggestions for overcoming resistance associated with the implementation of cpKPIs and provided insights into the future of pharmacy practice. The identification of barriers and facilitators to cpKPI implementation will be used to inform the implementation process on a local and national level.

Background: Evidence suggests that the underreporting of medication errors and near misses, collectively referred to as medication incidents (MIs), in the community pharmacy setting, is high. Despite the obvious negative implications, MIs present opportunities for pharmacy staff and regulatory authorities to learn from these mistakes and take steps to reduce the likelihood that they reoccur. However, these activities can only take place if such errors are reported and openly discussed., Objectives: This research proposes a model of factors influencing the reporting, service recovery, and organizational learning resulting from MIs within Canadian community pharmacies., Methods: The conceptual model is based on a synthesis of the literature and findings from a pilot study conducted among pharmacy management, pharmacists, and pharmacy technicians from 13 community pharmacies in Nova Scotia, Canada. The purpose of the pilot study was to identify various actions that should be taken to improve MI reporting and included staff perceptions of the strengths and weaknesses of their current MI-reporting process, desired characteristics of a new process, and broader external and internal activities that would likely improve reporting. Out of the 109 surveys sent, 72 usable surveys were returned (66.1% response rate). Multivariate analysis of variance found no significant differences among staff type in their perceptions of the current or new desired system but were found for broader initiatives to improve MI reporting. These findings were used for a proposed structural equation model (SEM)., Results: The SEM proposes that individual-perceived self-efficacy, MI process capability, MI process support, organizational culture, management support, and regulatory authority all influence the completeness of MI reporting, which, in turn, influences MI service recovery and learning., Conclusions: This model may eventually be used to enable pharmacy managers to make better decisions. By identifying risk factors that contribute to low MI reporting, recovery, and learning, it will be possible for regulators to focus their efforts on high-risk sectors and begin to undertake preventative educational interventions rather than relying solely on remedial activities., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Previous studies in animal models have suggested that bone marrow-derived interleukin-2 (IL-2) activated natural killer (A-NK*) cells can be used to eliminate residual leukemic cells both in vivo and in vitro. As part of initial studies to develop a clinical protocol to exploit this phenomenon in combination with culture purging of leukemic cells, we examined a number of variables that might affect either IL-2 stimulation of natural killer (NK) cell activity or maintenance of primitive hematopoietic cells in cultures suitable for manipulating human marrow autografts. Cytotoxicity of IL-2 A-NK cells was determined using K562 and Daudi target cells in a standard 4-hour 51Cr release assay. Primitive hematopoietic cells were quantitated using both direct clonogenic progenitor assays and the long-term culture-initiating cell (LTC-IC) assay. The latter measures a very primitive cell type that gives rise to clonogenic cells after > 5 weeks of culture on pre-established marrow feeder layers. Light-density (< 1.070 g/cm3) human marrow cells cultured at 10(6) cells/mL for 7 days at 37 degrees C in the presence of 1000 U of either natural or recombinant human IL-2 showed a marked generation of A-NK cell function that was not seen if IL-2 was not added. The cytotoxicity was the same whether the cells had been maintained in standard fetal calf serum (FCS)-supplemented medium or in LTC medium (which also contains horse serum and 10(-6) M hydrocortisone) or whether adherence of cells during the culture period was promoted or prevented. The level of NK activity in IL-2-stimulated cultures of marrow cells from patients with acute myeloid leukemia (AML) in remission was not significantly different from that obtained with normal marrow. The initial numbers of clonogenic cells in these samples were close to normal values (60%). For both normal and AML patient samples, these declined slightly during the 7-day culture period, regardless of whether IL-2 was present. Starting values for LTC-IC in patient marrow samples were markedly reduced to approximately 7% of normal values, but as in normal cultures, did not decrease further in culture with or without IL-2. These results indicate that activation of bone marrow-derived NK cells can be obtained in 7-day cultures containing IL-2 under conditions that preserve the most primitive hematopoietic cells currently detectable.

Previously we have described the isolation and characterization of a T-suppressor factor (TsF) from a T cell hybridoma (A10F), which has a degree of specificity for the DBA/2 mastocytoma P815. Administration of A10F intravenously at the time of tumor cell injection resulted in an accelerated rate of tumor growth, decreased cytotoxic T lymphocyte antitumor activity, and reduced survival time. In the work reported here, we have shown that administration of affinity-enriched A10F 7-14 days prior to tumor cell injection causes what appear to be reverse effects, in that an enhanced resistance to the P815 tumor is observed in vivo, an effect which we can correlate with the demonstration of antitumor cytotoxic T lymphocyte activity in vitro. These effects are dose-dependent since only doses of TsF at 20 micrograms or greater are effective. A similar effect was found when A10F was administered to DBA/2 mice 10 days prior to challenge with two unrelated tumors (L1210 and M-1). However, when another TsF (Fd11F) with apparent specificity for a nominal antigen was tested in this system, it had no effect on tumor growth.