Ultrasensitive NGS MRD assessment in Ph+ ALL: Prognostic impact and correlation with RT-PCR for BCR::ABL1.

Reverse transcription polymerase chain reaction (RT-PCR) for BCR::ABL1 is the most common and widely accepted method of measurable residual disease (MRD) assessment in Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL); however, RT-PCR may not be an optimal measure of MRD in many cases of Ph+ ALL. We evaluated the clinical impact of a highly sensitive next-generation sequencing (NGS) MRD assay (sensitivity of 10-6 ) and its correlation with RT-PCR for BCR::ABL1 in patients with Ph+ ALL. Overall, 32% of patients had a discordance between MRD assessment by RT-PCR and NGS, and 31% of patients who achieved NGS MRD negativity were PCR+ at the same timepoint. Among eight patients with long-term detectable BCR::ABL1 by PCR, six were PCR+/NGS-. These patients generally had stable PCR levels that persisted despite therapeutic interventions, and none subsequently relapsed; in contrast, patients who were PCR+/NGS+ had more variable PCR values that responded to therapeutic intervention. In a separate cohort of prospectively collected clinical samples, 11 of 65 patients (17%) with Ph+ ALL who achieved NGS MRD negativity had detectable BCR::ABL1 by PCR, and none of these patients relapsed. Relapse-free survival and overall survival were similar in patients who were PCR+/NGS- and PCR-/NGS-, suggesting that PCR for BCR::ABL1 did not provide additional prognostic information in patients who achieved NGS MRD negativity. NGS-based assessment of MRD is prognostic in Ph+ ALL and identifies patients with low-level detectable BCR::ABL1 who are unlikely to relapse nor to benefit from therapeutic interventions.

Clinical Management of Interproximal and Occlusal Caries in Children and Adolescents by Canadian Dentists: A Survey.

PURPOSE: Early restorative interventions may have important implications in young patients, and the International Caries Classification and Management System strongly recommends non-surgical strategies in the management of dental caries. We aimed to assess management of interproximal and occlusal caries in children and adolescents (≤18 years of age) by Canadian dentists. METHODS: An electronic survey was created and sent to members of Canadian provincial regulatory dental bodies. The survey included 11 questions on demographic factors and 3 clinical situations on dental caries management. RESULTS: The response rate was 4.6% (n = 702). To treat interproximal carious lesions limited to enamel, 442 dentists (63.0%) reported using surgical caries removal on a permanent molar and 502 dentists (71.5%) did the same for a primary tooth. For occlusal carious lesions, the corresponding numbers were 300 dentists (42.7%) for a permanent molar and 269 (38.3%) for a primary molar. Age, year of graduation and province of practice appear to have a significant impact on the restorative threshold. CONCLUSIONS: According to the latest evidence-based recommendations for caries management, the presence of cavitated enamel should be the main indication to restore, and non-surgical interventions for non-cavitated lesions confined to enamel should be prioritized. Results show that a good proportion of respondents have a lower restorative threshold, particularly for interproximal lesions.

Phase 2 study of lenalidomide maintenance for patients with high-risk acute myeloid leukemia in remission.

BACKGROUND: New drug combinations have led to significant improvements in remission rates for patients with acute myeloid leukemia (AML). However, many patients with high-risk AML who respond to their initial treatment and are not candidates for allogeneic stem cell transplantation (ASCT) will eventually relapse with poor outcomes. METHODS: In this phase 2 trial, the efficacy of lenalidomide maintenance was evaluated in patients with high-risk AML who had achieved their first or second remission after induction chemotherapy and at least 1 consolidation cycle and who were not candidates for immediate ASCT. Lenalidomide was given orally at 10 to 20 mg daily on days 1 to 28 of a 28-day cycle for up to 24 cycles. RESULTS: A total of 28 patients were enrolled in this study with a median age of 61 years (range, 24-87 years). The median number of cycles was 8 (range, 1-24 cycles). Ten patients (36%) completed 24 months of maintenance treatment. With a median follow-up of 22.5 months (range, 2.6-55 months), 12 patients (43%) relapsed after a median of 3 months (range, 0.7-23 months). The median duration of remission for all patients was 18.7 months (range, 0.7-55.1 months). The 2-year overall survival and relapse-free survival rates from the time of enrollment were 63% and 50%, respectively. Overall, lenalidomide was well tolerated; serious adverse events of grade 3 or 4, including rash (n = 5), thrombocytopenia (n = 4), neutropenia (n = 4), and fatigue (n = 2), were observed in 13 patients (46%). CONCLUSIONS: Lenalidomide is a safe and feasible maintenance strategy in patients with high-risk AML who are not candidates for ASCT, and it has beneficial effects for patients with negative measurable residual disease.

Factors influencing the engagement of older adults with neurocognitive disorders in the design of decision aids: A qualitative descriptive study.

OBJECTIVE: To identify factors influencing the engagement of older adults with neurocognitive disorders (NCDs) in the design of decision aids (DAs). METHODS: We conducted a qualitative descriptive study using semi-structured interviews with 23 older adults with NCDs who were accompanied by 27 caregivers. This is a secondary analysis of a published study to identify the features of DAs designed for this population and their caregivers. RESULTS: Several behaviours and attitudes of caregivers and researchers hindered the older adults' engagement in the DA design process. Specific communication strategies can be employed to support their engagement and overcome the communication challenges inherent to this population, such as memory, attention, hearing, or visual impairments. Adopting the appropriate attitude, taking the time, and providing guidance to the older person can help them focus on the topic, while developing trust between participants is a facilitator to obtain their feedback. CONCLUSION: Findings from this project could serve to inform the communication and co-design of DAs with older people with NCDs and their caregivers. PRACTICE IMPLICATIONS: Caregivers and researchers have key roles to play in facilitating communication with older persons with NCDs so they are empowered to help in co-designing DAs.

Development of a Competency Assessment Framework for Pharmacy Residents Participating in High-Fidelity Simulation and Creation of the SIMPHARM Assessment Tool.

Background: Simulations are used to improve professional practice across multiple health professions; however, the application of simulations in pharmacy curricula has been limited by a lack of evidence. Objectives: To delineate the competencies of pharmacy residents needed to assess their progress while participating in a high-fidelity simulation training program and to create a reliable assessment tool based on these competencies. Methods: A literature review was conducted, followed by application of a modified Delphi method. An assessment tool based on the results of these activities was drafted (in French). A second modified Delphi process was carried out to assess the reliability of the tool, and the tool was modified accordingly. Results: The literature search yielded a total of 2670 articles, of which 289 were included for analysis. The first modified Delphi process involved 19 experts in the first round and 10 experts in the second round. The Cronbach α was 0.866 (95% confidence interval [CI] 0.713-0.960), indicating good reliability. A total of 7 competencies were retained for inclusion in the SIMPHARM assessment tool: professionalism, leadership, teamwork, communication, critical thinking, preparation and packaging of medications, and pharmaceutical calculations. The second modified Delphi process involved 9 experts in the first round and 11 experts in the second round. The final Cronbach α was 0.877 (95% CI 0.741-0.960). Conclusions: To the authors' knowledge, this was one of the largest studies exploring pharmacy competencies in the context of simulations. This work yielded a reliable framework for the assessment of pharmacy residents' competencies. This assessment tool may help evaluators in assessing the competencies of pharmacy trainees after simulation training.

Contexte: Les simulations sont utilisées dans le but d'améliorer la pratique professionnelle de plusieurs professions de la santé; cependant, le manque de données probantes limite leur application dans les programmes d'enseignement en pharmacie. Objectifs: Décrire les compétences nécessaires pour évaluer le progrès des résidents en pharmacie qui participent à une formation basée sur un programme de simulation haute-fidélité; et mettre au point un outil d'évaluation fiable qui se base sur ces compétences. Méthodologie: Une revue de littérature a été effectuée, avant d'appliquer une méthode Delphi modifiée et de faire l'ébauche d'un outil d'évaluation (en français) basé sur les résultats de ces activités. Une deuxième méthode Delphi modifiée a été appliquée pour évaluer la fiabilité de l'outil et celui-ci a fait l'objet de modifications en conséquence. Résultats: La revue de littérature a donné un total de 2670 articles, parmi lesquels 289 ont été inclus à des fins d'analyse. La première méthode Delphi modifiée a impliqué 19 experts pour la première phase, et 10 pour la seconde. Le coefficient alpha de Cronbach était de 0,866 (intervalle de confiance [IC] à 95 % 0,713­0,960), indiquant une bonne fiabilité. Au total, 7 compétences à inclure dans l'outil d'évaluation ont été retenues: professionnalisme, leadership, travail d'équipe, communication, pensée critique, préparation et conditionnement des médicaments, et calculs pharmaceutiques. La deuxième méthode Delphi modifiée impliquait quant à elle 9 experts pour la première phase et 11 pour la seconde. Le coefficient alpha de Cronbach final était de 0,877 (IC à 95 % 0,741­0,960). Conclusions: À la connaissance des auteurs, cette étude était l'une des plus importantes se penchant sur les compétences en pharmacie dans le contexte des simulations. Ce travail a donné lieu à un cadre de référence fiable pour évaluer les compétences des résidents en pharmacie. Cet outil d'évaluation pourrait aider les évaluateurs à évaluer les compétences des stagiaires à la suite d'une formation par simulation.

Fast and high-resolution ion chromatography at high pH on short columns packed with 1.8 microm surfactant coated silica reverse-phase particles.

Rapid ion chromatographic separations of small inorganic anions are performed on columns packed with high-pH resistant Zorbax Extend-C18 1.8 microm silica particles. Seven anions (iodate, chloride, nitrite, bromide, nitrate, phosphate, sulphate) are separated with 1.3 and 2 cm long x 0.46 cm I.D. C18 columns coated with the surfactant didodecyldimethylammonium bromide (DDAB). A 40 s separation is achieved at 2 mL/min with a 2.5 mM 4-hydroxybenzoic acid eluent at pH 10. Finally, the DDAB removal procedure is improved to eliminate the pressure build-up caused by precipitation of the surfactant in the column upon uncoating.

Achieving rapid low-pressure ion chromatography separations on short silica-based monolithic columns.

Short silica-based monolithic columns (0.5-1 cm) are coated with the surfactant didodecyldimethylammonium bromide (DDAB) and used for fast ion exchange separations of small inorganic anions. Sources of extra-column band broadening are assessed and minimized to obtain separations of seven analytes (iodate, chloride, nitrate, bromide, nitrite, phosphate, sulphate) in two minutes at 2 mL/min. Eluents used are 6 and 9 mM 4-cyanophenol at pH 7.3-7.4 or 5 mM 4-hydroxybenzoic acid at pH 5.6. DDAB coating stability is improved by 15-fold by the addition of a DDAB coated pre-column before the injection valve. Separations are obtained using a low-pressure glass syringe.

Characterization of Two Dinoflagellate Cold Shock Domain Proteins.

Roughly two-thirds of the proteins annotated as transcription factors in dinoflagellate transcriptomes are cold shock domain-containing proteins (CSPs), an uncommon condition in eukaryotic organisms. However, no functional analysis has ever been reported for a dinoflagellate CSP, and so it is not known if they do in fact act as transcription factors. We describe here some of the properties of two CSPs from the dinoflagellate Lingulodinium polyedrum, LpCSP1 and LpCSP2, which contain a glycine-rich C-terminal domain and an N-terminal cold shock domain phylogenetically related to those in bacteria. However, neither of the two LpCSPs act like the bacterial CSP, since they do not functionally complement the Escherichia coli quadruple cold shock domain protein mutant BX04, and cold shock does not induce LpCSP1 and LpCSP2 to detectable levels, based on two-dimensional gel electrophoresis. Both CSPs bind to RNA and single-stranded DNA in a nonspecific manner in electrophoretic mobility shift assays, and both proteins also bind double-stranded DNA nonspecifically, albeit more weakly. These CSPs are thus unlikely to act alone as sequence-specific transcription factors. IMPORTANCE Dinoflagellate transcriptomes contain cold shock domain proteins as the major component of the proteins annotated as transcription factors. We show here that the major family of cold shock domain proteins in the dinoflagellate Lingulodinium do not bind specific sequences, suggesting that transcriptional control is not a predominant mechanism for regulating gene expression in this group of protists.

High-performance liquid chromatographic separation and indirect fluorescence detection of thiols.

A fluorescent post-column reaction detection scheme has been devised for selective determination of thiols. The post-column reagent is 40 microM Cd2+ and 100 microM 8-hydroxyquinoline-5-sulfonic acid (HQS) in non-complexing buffer at pH 10. HQS complexes Cd2+ to form a fluorescent product. Thiols in the HPLC effluent compete for complexation of the Cd2+, resulting in a decrease in the fluorescence response. Detection limits of 0.2 microM (0.04 ppm) are achieved for cysteine, homocysteine and glutathione in a 5 min separation. Recoveries from spiked synthetic urine samples are 87-120%.

Effects of stimulus rate and gender on the auditory middle latency response.

The effects of stimulus rate and gender on the auditory middle latency response (AMLR) waveforms were examined in 20 young adult male and female subjects. Four different repetition rates were presented to subjects (1.1/sec, 4.1/sec, 7.7/ sec, and 11.3/sec). Stimulus repetition rate had a significant effect on Pa latency, Pa amplitude, and Pb amplitude. Pa and Pb amplitudes decreased with increasing the stimulus rate, and Pa latency significantly increased with increasing the stimulus rate. No significant differences were seen on Pb latency or site of recording. Gender had a significant effect on Pa latency and Pa amplitude. Pa latencies were longer in male subjects, and Pa amplitudes were larger in female subjects. Gender did not have a significant effect on the Pb waveform.

Target profiling of a small library of phosphodiesterase†5 (PDE5) inhibitors using chemical proteomics.

Inhibitors of phosphodiesterase 5 (PDE5) are widely used for the treatment of erectile dysfunction and pulmonary hypertension. The commercially available inhibitors are effective, well-tolerated drugs, but differ in their phosphodiesterase specificity. To explore and manipulate the specificity of PDE5 inhibitors, a small library of four inhibitors was synthesized using the structure of known PDE5 inhibitors as a scaffold. Their inhibitory potency towards PDE5 and related family members was evaluated. Next, they were immobilized on a matrix to perform affinity pull-down assays in rat testis tissue, followed by mass spectrometric (MS) analysis. By using unique peptide spectral counts of identified proteins in the MS analysis, we were able to assess the relative binding of these inhibitors to a large set of proteins, allowing the determination of their selectivity profiles in vitro. For selected proteins of interest, the results were verified using quantitative isotopic dimethyl labeling and immunoblotting, and isothermal titration calorimetry (ITC). For the PDE5 inhibitors, our data reveal that even slight chemical modifications can bias their selectivity significantly towards other interacting proteins, opening up the potential of these compounds to be used as scaffolds for the development of inhibitors for new protein targets. In a broad sense, we demonstrate that the combination of chemical proteomics and unique peptide spectral counting allows for the confident and facile analysis of the differential interactome of bioactive small molecules.

Quantifying cross-tissue diversity in proteasome complexes by mass spectrometry.

The composition of 20S mouse proteasome complexes isolated from mice heart, kidney, liver, lung, thymus and spleen was compared using quantitative mass spectrometry. Significant variety was observed in hybrid classes of immunoproteasomes which may have implications for the use of proteasome targeted inhibitors.

HPLC simultaneous analysis of thiols and disulfides: on-line reduction and indirect fluorescence detection without derivatization.

A liquid chromatography method with indirect fluorescence detection has been developed for simultaneous detection of cysteine, cystine, homocysteine, homocystine, glutathione and glutathione disulfide. After separation in their native forms, a post-column solution of tris(2-carboxyethyl)phosphine (TCEP) at 32 mM, pH 8 is added to reduce the disulfides on-line to the corresponding thiol. The effluent is then merged with a second post-column solution of the highly fluorescent complex Cd(HQS)(2)(2-). The cadmium is complexed by the eluting thiols, effectively quenching the fluorescence. Optimization of the separation, the on-line reduction and the indirect fluorescence detection are discussed. Detection limits from 0.3-4.3 microM (0.04 to 2.6 ppm) are achieved for the six analytes in a 20 min separation.