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Purpose of program: Operative wait times for non-oncology-related procedures continue to rise in Canada, and this was further exacerbated by the COVID-19 pandemic. These challenges will remain prevalent beyond the pandemic given the limited number of acute care beds and resources required to care for patients. As a result, the need for innovative approaches to optimize the utilization of health care resources while maintaining equitable and timely access is needed. In this report, we describe the development of a collaborative ambulatory parathyroidectomy program between two centers in Toronto, allowing for more expedient surgical treatment of secondary hyperparathyroidism among patients from a large dialysis program. Sources of information: The need for expanded access to surgical care for secondary hyperparathyroidism was identified through interdepartmental communication between referring nephrologists and surgeons at Sunnybrook Health Sciences Centre and Women’s College Hospital, respectively. Methods: A multidisciplinary ambulatory parathyroidectomy planning team was formed that included nephrologists, endocrine surgeons, nurses, and patient care managers to conduct a needs assessment. It was identified that patients had long wait times, and to address that gap in care, a protocol was developed to identify suitable patients requiring treatment. The teams created a plan to coordinate patient care and transfers. A clinical tool and protocol for post-operative management of hypocalcemia was developed using a Delphi model, gathering input from many members of the care team. The Delphi process to finalize the protocol included a series of virtual meetings over a period of about 4 months with various stakeholders and included input from two departmental heads (medicine and surgery), three nephrologists, a nurse practitioner, a patient care manager, and two nurse educators. Meetings involved core members of the Nephrology Quality Improvement and Patient Safety at Sunnybrook Health Sciences Centre and finalized protocol was agreed upon by members of this group at a quarterly meeting. Key findings: In this article, we describe the development, initial deployment, and planned assessment of the ambulatory parathyroidectomy program at the Women’s College Hospital and Sunnybrook Health Sciences Centre. The primary aim of the program is to increase accessibility to parathyroidectomy for secondary hyperparathyroidism. A secondary aim was to allow patients to have streamlined care with a team that is well versed with maintenance dialysis needs and optimizing treatment of post operative hypocalcemia through standardized protocols. Limitations: Ambulatory parathyroidectomy relies on effective communication, flow, and availability of acute care beds. It is anticipated that occasionally, unexpected hospital demands, and health care disruptions may occur, which can limit efficiency of the program. We will also need to examine the cost-effectiveness of this program as it may improve access but increase costs related to the procedure. As the program is implemented, useful adaptations and policies to our protocol to help mitigate these limitations will be documented and published in our outcomes report. Implications: Ontario residents with chronic kidney disease with secondary hyperparathyroidism who have failed medical management will have increased and timely access to parathyroidectomy.

Introduction Heart failure (HF) symptoms improve through self-care, for which adherence remains low among patients despite the provision of education for these behaviours by clinical teams. Open Access Digital Community Promoting Self-Care, Peer Support and Health Literacy (ODYSSEE–vCHAT) combines automated digital counselling with social network support to improve mortality and morbidity, engagement with self–care materials, and health-related quality of life.Methods and analysis Use of ODYSSEE-vCHAT via Internet-connected personal computer by 162 HF patients will be compared with a control condition over 22 months. The primary outcome is a composite index score of all-cause mortality, all-cause emergency department visits, and HF-related hospitalisation at trial completion. Secondary outcomes include individual components of the composite index, engagement with self-care materials, and patient-reported measures of physical and psychosocial well-being, disease management, health literacy, and substance use. Patients are recruited from tertiary care hospitals in Toronto, Canada and randomised on a 1:1 ratio to both arms of the trial. Online assessments occur at baseline (t=0), months 4, 8 and 12, and trial completion. Ordinal logistic regression analyses and generalised linear models will evaluate primary and secondary outcomes.Ethics and dissemination The trial has been approved by the research ethics boards at the University Health Network (20-5960), Sunnybrook Hospital (5117), and Mount Sinai Hospital (21-022-E). Informed consent of eligible patients occurs in person or online. Findings will be shared with key stakeholders and the public. Results will allow for the preparation of a Canada-wide phase III trial to evaluate the efficacy of ODYSSEE-vCHAT in improving clinical outcomes and raising the standard of outpatient care.Trial registration number NCT04966104

Objectives Home haemodialysis (HD) and peritoneal dialysis (PD) have seen growth in utilisation around the globe over the last few years. However, home dialysis, with its attendant technical complexity and risk of adverse events continues to pose challenges for wider adoption. We examined whether differences in patients’ learning styles are associated with differing risk of adverse events in both home HD and PD patients.Design Retrospective cohort study.Setting Tertiary care hospital in Toronto, Ontario, Canada.Participants One hundred and eighteen prevalent adult (≥18 years) home dialysis patients (40 PD and 78 home HD) were enrolled. Patients on home dialysis for less than 6 months or receiving home nursing assistance for dialysis were excluded from the study.Interventions Enrolled patients completed (VARK) Visual, Aural, Reading-writing and Kinesthetic questionnaires to determine learning styles.Primary and secondary outcome measures Home HD and PD adverse events were identified within 6 months of completing home dialysis training. Event rates were then stratified and compared according to learning styles.Results Thirty patients had a total of 53 adverse events. We used logistic regression analysis to determine unadjusted and adjusted ORs for a single adverse event. Non-visual learners were 4.35 times more likely to have an adverse event (p=0.001). After adjusting for age, gender, dialysis modality, training duration, dialysis vintage, prior renal replacement therapy, visual impairment, education and literacy, an adverse event was still four times more likely among non-visual learners compared to visual learners (p=0.008). A subgroup analysis of home HD patients showed adverse events were more likely among non-visual learners (OR 11.1; p=0.003), whereas PD patients showed a trend for more adverse events in non-visual learners (OR: 1.60; p=0.694).Conclusions Different learning styles in home dialysis patients exist. Visual learning styles are associated with fewer adverse events in home dialysis patients within the first 6 months of completing training. Individualisation of home dialysis training by learning style is warranted.

When some perceptual-motor relationships are reversed, participants might adopt a global inhibition strategy that replaces all normal movements with reversed movements. In two experiments, participants practiced moving a cursor from a start position to target locations. In a perceptual-motor reversal condition, in which horizontal but not vertical movements were reversed, participants were trained to move only to certain locations. Testing involved moving to all locations under the same reversal condition. Training on a subset of locations yielded partial transfer to untrained locations. These results support a global inhibition hypothesis modified to include both midcourse corrective movements and training specificity. [ABSTRACT FROM AUTHOR]

In two experiments, we demonstrated two types of strategies (rule-based and memory-based) and strategy transitions within the same binary classification task. The strategy that dominated later in practice depended on the difficulty of the operative classification rule and on the salience of the cue for that rule. Accuracy increased over practice trials, and response times were faster for the dominant strategy, either rule or memory. Rule retention was superior to stimulus item retention, so that, even for participants who preferred a memory-based strategy, a rule-based strategy dominated at least temporarily after a 1-week interval. Strategy use over trials and the retention interval reflected a given task's affordance of a shift between rule- and memory-based processes. [ABSTRACT FROM AUTHOR]

In two experiments, we examined training, retention, and transfer of a duration production skill in a prospective paradigm. Participants were trained with feedback and then were either tested immediately for transfer without feedback or retrained with feedback 1 week later. There were three training and retraining conditions, two involving secondary tasks. Retention of the duration production skill was perfect across the 1-week delay when the secondary task condition was unchanged, but there was no skill transfer when that condition was changed. These findings demonstrate specificity of training, with the assumption that the cognitive operations learned during duration production training incorporate requirements of the secondary task. More generally, this study challenges the current practice in which training conditions often do not match eventual testing conditions. [ABSTRACT FROM AUTHOR]

Background: Prenatal alcohol exposure (PAE) can result in lifelong disabilities known as foetal alcohol spectrum disorder (FASD) and is associated with childhood growth deficiencies and increased bone fracture risk. However, the effects of PAE on the adult skeleton remain unclear and any potential sexual dimorphism is undetermined. Therefore, we utilised a murine model to examine sex differences with PAE on in vitro bone formation, and in the juvenile and adult skeleton., Methods: Pregnant C57BL/6J female mice received 5% ethanol in their drinking water during gestation. Primary calvarial osteoblasts were isolated from neonatal offspring and mineralised bone nodule formation and gene expression assessed. Skeletal phenotyping of 4- and 12-week-old male and female offspring was conducted by micro-computed tomography (µCT), 3-point bending, growth plate analyses, and histology., Results: Osteoblasts from male and female PAE mice displayed reduced bone formation, compared to control (≤ 30%). Vegfa, Vegfb, Bmp6, Tgfbr1, Flt1 and Ahsg were downregulated in PAE male osteoblasts only, whilst Ahsg was upregulated in PAE females. In 12-week-old mice, µCT analysis revealed a sex and exposure interaction across several trabecular bone parameters. PAE was detrimental to the trabecular compartment in male mice compared to control, yet PAE females were unaffected. Both male and female mice had significant reductions in cortical parameters with PAE. Whilst male mice were negatively affected along the tibial length, females were only distally affected. Posterior cortical porosity was increased in PAE females only. Mechanical testing revealed PAE males had significantly reduced bone stiffness compared to controls; maximum load and yield were reduced in both sexes. PAE had no effect on total body weight or tibial bone length in either sex. However, total growth plate width in male PAE mice compared to control was reduced, whilst female PAE mice were unaffected. 4-week-old mice did not display the altered skeletal phenotype with PAE observed in 12-week-old animals., Conclusions: Evidence herein suggests, for the first time, that PAE exerts divergent sex effects on the skeleton, possibly influenced by underlying sex-specific transcriptional mechanisms of osteoblasts. Establishing these sex differences will support future policies and clinical management of FASD., (© 2024. The Author(s).)

Introduction: Chondrocytes are continuously exposed to loads placed upon them. Physiological loads are pivotal to the maintenance of articular cartilage health, while abnormal loads contribute to pathological joint degradation. Similarly, the growth plate cartilage is subject to various loads during growth and development. Due to the high-water content of cartilage, hydrostatic pressure is considered one of the main biomechanical influencers on chondrocytes and has been shown to play an important role in the mechano-regulation of cartilage., Methods: Herein, we conducted RNAseq analysis of ex vivo hip cap (articular), and metatarsal (growth plate) cartilage cultures subjected to physiological (5 MPa) and injurious (50 MPa) hydrostatic pressure, using the Illumina platform (n = 4 replicates)., Results: Several hundreds of genes were shown to be differentially modulated by hydrostatic pressure, with the majority of these changes evidenced in hip cap cartilage cultures (375 significantly upregulated and 322 downregulated in 5 MPa versus control; 1022 upregulated and 724 downregulated in 50 MPa versus control). Conversely, fewer genes were differentially affected by hydrostatic pressure in the metatarsal cultures (5 significantly upregulated and 23 downregulated in 5 MPa versus control; 7 significantly upregulated and 19 downregulated in 50 MPa versus control). Using Gene Ontology annotations for Biological Processes, in the hip cap data we identified a number of pathways that were modulated by both physiological and injurious hydrostatic pressure. Pathways upregulated in response to 50 MPa versus control, included those involved in the generation of precursor metabolites and cellular respiration. Biological processes that were downregulated in this tissue included ossification, connective tissue development, and chondrocyte differentiation., Discussion: Collectively our data highlights the divergent chondrocyte phenotypes in articular and growth plate cartilage. Further, we show that the magnitude of hydrostatic pressure application has distinct effects on gene expression and biological processes in hip cap cartilage explants. Finally, we identified differential expression of a number of genes that have previously been identified as osteoarthritis risk genes, including Ctsk, and Chadl. Together these data may provide potential genetic targets for future investigations in osteoarthritis research and novel therapeutics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Bourne, Hesketh, Sharma, Bucca, Bush and Staines.)

Extracellular pyrophosphate (PP i ) is well known for its fundamental role as a physiochemical mineralisation inhibitor. However, information about its direct actions on bone cells remains limited. This study shows that PP i decreased osteoclast formation and resorptive activity by ≤50 %. These inhibitory actions were associated with reduced expression of genes involved in osteoclastogenesis (Tnfrsf11a, Dcstamp) and bone resorption (Ctsk, Car2, Acp5). In osteoblasts, PP i present for the entire (0-21 days) or latter stages of culture (7-21/14-21 days) decreased bone mineralisation by ≤95 %. However, PP i present for the differentiation phase only (0-7/0-14 days) increased bone formation (≤70 %). Prolonged treatment with PP i resulted in earlier matrix deposition and increased soluble collagen levels (≤2.3-fold). Expression of osteoblast (RUNX2, Bglap) and early osteocyte (E11, Dmp1) genes along with mineralisation inhibitors (Spp1, Mgp) was increased by PP i (≤3-fold). PP i levels are regulated by tissue non-specific alkaline phosphatase (TNAP) and ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1). PP i reduced NPP1 expression in both cell types whereas TNAP expression (≤2.5-fold) and activity (≤35 %) were increased in osteoblasts. Breakdown of extracellular ATP by NPP1 represents a key source of PP i . ATP release from osteoclasts and osteoblasts was decreased ≤60 % by PP i and by a selective TNAP inhibitor (CAS496014-12-2). Pertussis toxin, which prevents Gα i subunit activation, was used to investigate whether G-protein coupled receptor (GPCR) signalling mediates the effects of PP i . The actions of PP i on bone mineralisation, collagen production, ATP release, gene/protein expression and osteoclast formation were abolished or attenuated by pertussis toxin. Together these findings show that PP i , modulates differentiation, function and gene expression in osteoblasts and osteoclasts. The ability of PP i to alter ATP release and NPP1/TNAP expression and activity indicates that cells can detect PP i levels and respond accordingly. Our data also raise the possibility that some actions of PP i on bone cells could be mediated by a Gα i -linked GPCR., Competing Interests: Declaration of competing interest The authors have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Bone cells are known to express multiple P2 receptor subtypes, and the functional effects of receptor activation have been described for many of these. One exception is the P2X4 receptor, which despite strong expression in osteoblasts and osteoclasts, has no defined functional activity. This study used the selective P2X4 receptor antagonists, 5-BDBD and PSB-12062, to investigate the role of this receptor in bone. Both antagonists (≥ 0.1 μM) dose-dependently decreased bone formation by 60-100%. This was accompanied by a ≤ 70% decrease in alkaline phosphatase activity, a ≤ 40% reduction in cell number, and a ≤ 80% increase in the number of adipocytes present in the culture. The analysis of gene expression showed that levels of osteoblast marker genes (e.g. Alpl, Bglap) were decreased in 5-BDBD treated cells. Conversely, expression of the adipogenic transcription factor PPARG was increased 10-fold. In osteoclasts, high doses of both antagonists were associated with a reduction in osteoclast formation and resorptive activity by ≤ 95% and ≤ 90%, respectively. Taken together, these data suggest that the P2X4 receptor plays a role in modulating bone cell function. In particular, it appears to influence osteoblast differentiation favouring the osteogenic lineage over the adipogenic lineage., (© 2022. The Author(s).)

Arterial medial calcification (AMC) is the deposition of calcium phosphate in the arteries. AMC is widely thought to share similarities with physiological bone formation; however, emerging evidence suggests several key differences between these processes. N-acetylcysteine (NAC) displays antioxidant properties and can generate hydrogen sulphide (H 2 S) and glutathione (GSH) from its deacetylation to l-cysteine. This study found that NAC exerts divergent effects in vitro, increasing osteoblast differentiation and bone formation by up to 5.5-fold but reducing vascular smooth muscle cell (VSMC) calcification and cell death by up to 80%. In vivo, NAC reduced AMC in a site-specific manner by 25% but had no effect on the bone. The actions of l-cysteine and H 2 S mimicked those of NAC; however, the effects of H 2 S were much less efficacious than NAC and l-cysteine. Pharmacological inhibition of H 2 S-generating enzymes did not alter the actions of NAC or l-cysteine; endogenous production of H 2 S was also unaffected. In contrast, NAC and l-cysteine increased GSH levels in calcifying VSMCs and osteoblasts by up to 3-fold. This suggests that the beneficial actions of NAC are likely to be mediated via the breakdown of l-cysteine and the subsequent GSH generation. Together, these data show that while the molecular mechanisms driving the actions of NAC appear similar, the downstream effects on cell function differ significantly between osteoblasts and calcifying VSMCs. The ability of NAC to exert these differential actions further supports the notion that there are differences between the development of pathological AMC and physiological bone formation. NAC could represent a therapeutic option for treating AMC without exerting negative effects on bone., (© 2021 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.)

Arterial medial calcification (AMC), the deposition of hydroxyapatite in the medial layer of the arteries, is a known risk factor for cardiovascular events. Oxidative stress is a known inducer of AMC and endogenous antioxidants, such as glutathione (GSH), may prevent calcification. GSH synthesis, however, can be limited by cysteine levels. Therefore, we assessed the effects of the cysteine prodrug 2-oxothiazolidine-4-carboxylic acid (OTC), on vascular smooth muscle cell (VSMC) calcification to ascertain its therapeutic potential. Human aortic VSMCs were cultured in basal or mineralising medium (1 mM calcium chloride/sodium phosphate) and treated with OTC (1-5 mM) for 7 days. Cell-based assays and western blot analysis were performed to assess cell differentiation and function. OTC inhibited calcification ≤90%, which was associated with increased ectonucleotide pyrophosphatase/phosphodiesterase activity, and reduced apoptosis. In calcifying cells, OTC downregulated protein expression of osteoblast markers (Runt-related transcription factor 2 and osteopontin), while maintaining expression of VSMC markers (smooth muscle protein 22α and α-smooth muscle actin). GSH levels were significantly reduced by 90% in VSMCs cultured in calcifying conditions, which was associated with declines in expression of gamma-glutamylcysteine synthetase and GSH synthetase. Treatment of calcifying cells with OTC blocked the reduction in expression of both enzymes and prevented the decline in GSH. This study shows OTC to be a potent and effective inhibitor of in vitro VSMC calcification. It appears to maintain GSH synthesis which may, in turn, prevent apoptosis and VSMCs gaining osteoblast-like characteristics. These findings may be of clinical relevance and raise the possibility that treatment with OTC could benefit patients susceptible to AMC., (© 2020 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.)

Biomineralisation, the deposition of mineral onto a matrix, can be both a physiological and pathological process. Bone formation involves the secretion of an extracellular matrix (ECM) by osteoblasts and subsequent mineralisation of that matrix. It is regulated by a number of local and systemic factors and is necessary for maintenance of normal bone health. Conversely, mineralisation (or calcification) of soft tissues, including the vasculature, is detrimental to that tissue, leading to diseases such as arterial medial calcification (AMC). The mechanisms underlying AMC development are not fully defined, though it is thought that vascular smooth muscle cells (VSMCs) drive this complex, cell-mediated process. Similarly, AMC is regulated by a variety of enzymes and molecules, many of which have already been implicated in the regulation of bone mineralisation. This review will provide an overview of the similar, and sometimes opposing effects of these signalling molecules on the regulation of bone mineralisation and AMC.

Arterial medial calcification (AMC) has been associated with phenotypic changes in vascular smooth muscle cells (VSMCs) that reportedly makes them more osteoblast-like. Previous work has shown that ATP/UTP can inhibit AMC directly via P2 receptors and indirectly by NPP1-mediated hydrolysis to produce the mineralisation inhibitor, pyrophosphate (PP i ). This study investigated the role of P2X receptors in the inhibitory effects of extracellular nucleotides on VSMC calcification. We found that Bz-ATP, α,β-meATP and β,γ-meATP inhibited calcification by up to 100%. Culture in a high-phosphate medium (2 mM) was associated with increased VSMC death and apoptosis; treatment with Bz-ATP, α,β-meATP and β,γ-meATP reduced apoptosis to levels seen in non-calcifying cells. Calcification was also associated with alterations in the protein levels of VSMC (e.g. SM22α and SMA) and osteoblast-associated (e.g. Runx2 and osteopontin) markers; Bz-ATP, α,β-meATP and β,γ-meATP attenuated these changes in protein expression. Long-term culture with Bz-ATP, α,β-meATP and β,γ-meATP resulted in lower extracellular ATP levels and an increased rate of ATP breakdown. P2X receptor antagonists failed to prevent the inhibitory effects of these analogues suggesting that they act via P2X receptor-independent mechanisms. In agreement, the breakdown products of α,β-meATP and β,γ-meATP (α,β-meADP and methylene diphosphonate, respectively) also dose-dependently inhibited VSMC calcification. Furthermore, the actions of Bz-ATP, α,β-meATP and β,γ-meATP were unchanged in VSMCs isolated from NPP1-knockout mice, suggesting that the functional effects of these compounds do not involve NPP1-mediated generation of PP i . Together, these results indicate that the inhibitory effects of ATP analogues on VSMC calcification and apoptosis in vitro may be mediated, at least in part, by mechanisms that are independent of purinergic signalling and PP i .

Arterial medial calcification (AMC) is the deposition of calcium phosphate mineral, often as hydroxyapatite, in the medial layer of the arteries. AMC shares some similarities to skeletal mineralisation and has been associated with the transdifferentiation of vascular smooth muscle cells (VSMCs) towards an osteoblast-like phenotype. This study used primary mouse VSMCs and calvarial osteoblasts to directly compare the established and widely used in vitro models of AMC and bone formation. Significant differences were identified between osteoblasts and calcifying VSMCs. First, osteoblasts formed large mineralised bone nodules that were associated with widespread deposition of an extracellular collagenous matrix. In contrast, VSMCs formed small discrete regions of calcification that were not associated with collagen deposition and did not resemble bone. Second, calcifying VSMCs displayed a progressive reduction in cell viability over time (≤7-fold), with a 50% increase in apoptosis, whereas osteoblast and control VSMCs viability remained unchanged. Third, osteoblasts expressed high levels of alkaline phosphatase (TNAP) activity and TNAP inhibition reduced bone formation by to 90%. TNAP activity in calcifying VSMCs was ∼100-fold lower than that of bone-forming osteoblasts and cultures treated with β-glycerophosphate, a TNAP substrate, did not calcify. Furthermore, TNAP inhibition had no effect on VSMC calcification. Although, VSMC calcification was associated with increased mRNA expression of osteoblast-related genes (e.g. Runx2, osterix, osteocalcin, osteopontin), the relative expression of these genes was up to 40-fold lower in calcifying VSMCs versus bone-forming osteoblasts. In summary, calcifying VSMCs in vitro display some limited osteoblast-like characteristics but also differ in several key respects: 1) their inability to form collagen-containing bone; 2) their lack of reliance on TNAP to promote mineral deposition; and, 3) the deleterious effect of calcification on their viability., (Crown Copyright © 2019. Published by Elsevier Inc. All rights reserved.)

Theories of memory must account for memory performance during both the acquisition (i.e., ongoing learning) and retention (i.e., following disuse) stages of training. One factor affecting both stages is whether repeated encounters with a set of material occur with no delay between blocks (massed) or alternating with another intervening task (spaced). Whereas the retention advantage for spaced over massed practice is well accounted for by some current theories of memory, theories of decay or general interference predict massed, rather than spaced, advantages during acquisition. In a series of 3 experiments, we show that the effects of spacing on acquisition depend on the relationship between primary and delay tasks. Specifically, massed acquisition advantages occur only in the presence of code-specific interference (the engagement in two alternating tasks both emphasizing the same processing code, such as verbal or spatial processing codes; e.g., learning letter-number pairs and reading text), whereas spaced acquisition advantages are observed only when code-specific interference is absent. These results present a challenge for major theories of memory. Furthermore, we argue that code-specific interference is important for researchers of the spacing and interleaving effects to take into consideration, as the relationship between the alternating tasks used has a substantial impact on acquisition performance.

This chapter describes the isolation, culture, and staining of osteoblasts. The key advantages of this assay are that it allows direct measurement of bone matrix deposition and mineralization, as well as yielding good quantities of osteoblasts at defined stages of differentiation for molecular and histological analysis. An additional focus of this chapter will be the culture of osteoblasts from less conventional animal species.

Arterial medial calcification (AMC) is thought to share some outward similarities to skeletal mineralization and has been associated with the transdifferentiation of vascular smooth muscle cells (VSMCs) to an osteoblast-like phenotype. ATP and UTP have previously been shown to inhibit bone mineralization. This investigation compared the effects of extracellular nucleotides on calcification in VSMCs with those seen in osteoblasts. ATP, UTP and the ubiquitous mineralization inhibitor, pyrophosphate (PP i ), dose dependently inhibited VSMC calcification by ≤85%. Culture of VSMCs in calcifying conditions was associated with an increase in apoptosis; treatment with ATP, UTP, and PP i reduced apoptosis to levels seen in non-calcifying cells. Extracellular nucleotides had no effect on osteoblast viability. Basal alkaline phosphatase (TNAP) activity was over 100-fold higher in osteoblasts than VSMCs. ATP and UTP reduced osteoblast TNAP activity (≤50%) but stimulated VSMC TNAP activity (≤88%). The effects of extracellular nucleotides on VSMC calcification, cell viability and TNAP activity were unchanged by deletion or inhibition of the P2Y 2 receptor. Conversely, the actions of ATP/UTP on bone mineralization and TNAP activity were attenuated in osteoblasts lacking the P2Y 2 receptor. Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) hydrolyses ATP and UTP to produce PP i . In both VSMCs and osteoblasts, deletion of NPP1 blunted the inhibitory effects of extracellular nucleotides suggesting involvement of P2 receptor independent pathways. Our results show that although the overall functional effect of extracellular nucleotides on AMC and bone mineralization is similar there are clear differences in the cellular mechanisms mediating these actions., (© 2017 Wiley Periodicals, Inc.)

To study the relative merits of three training principles - difficulty of training, specificity of training, and variability of training - subjects were trained to follow navigation instructions to move in a grid on a computer screen. Subjects repeated and then followed the instructions by mouse clicking on the grid. They were trained, given a short distractor task, and then tested. There were three groups, each receiving different message lengths during training: easy (short lengths), hard (long lengths), and mixed (all lengths), with all subjects given all lengths at test. At test, the mixed group was best on most lengths, the easy group was better than the hard group on short lengths, and the hard group was better than the easy group on long lengths. The results support the advantages of both specificity and variability of training but do not support the hypothesis that difficult training of the form used here would lead to overall best performance at test.

Four sets of cognitive psychology experiments are aimed to identify and empirically support principles of training intended to increase training effectiveness. Each set supports a different training principle: strategic use of knowledge (learning and memory are facilitated whenever preexisting knowledge can be used as a mediator in the process of acquisition), training difficulty (conditions that cause desirable difficulties during learning might facilitate later retention and transfer), mental practice (mental practice can retard forgetting and promote transfer of training, sometimes to a larger extent than can physical practice), and cognitive antidote (adverse effects of prolonged work on routine tasks can be mitigated by adding cognitive complications). The first 2 principles are focused on knowledge acquisition and the last 2 on skill acquisition. Nevertheless, the principles converge on similar prescriptions: Contrary to intuition and common practice, the trainer is recommended to increase the complexity of the training situation rather than use the simplest methods available.

Two experiments examined transfer of a prospective, time production skill under conditions involving changes in concurrent task requirements. Positive transfer of the time production skill might be expected only when the attentional demands of the concurrent task were held constant from training to test. However, some positive transfer was found even when the concurrent task at retraining was made either easier or more difficult than the concurrent task learned during training. The amount and direction of transfer depended more on the pacing of the stimuli in the secondary task than on the difficulty of the secondary task, even though difficulty affects attentional demands more. These findings are consistent with the procedural reinstatement principle of skill learning, by which transfer from one task to another depends on an overlap in procedures required by the two skills.

Any technique that conserves classroom instructional time without sacrificing the amount learned is of great educational value. This research compared a laboratory analogue of the clicker technique to analogues of other classroom pedagogical methods that all involve repeated testing during teaching. The clicker analogue mimics the classroom practice of dropping material that is understood by the majority of the class, as revealed by testing with clicker questions, from further lecture. A fact learning and retrieval paradigm was used, in which college students learned facts about unfamiliar countries. Compressing instruction time based on group-level performance produced as much learning as no compression and as compression based on individual-level performance. Results suggest that the clicker technique is an efficient and cost-effective method of conserving instructional time without loss of amount learned.

In a continuous memory-updating paradigm, subjects studied name-color associations and were tested later for the color associate given the name. The default color response was made in one location, but on predesignated trials the response was required to be made in a special location. Memory for the color associated with a given name was assessed after short and long retention intervals when both default and special responses were required. Separate measures were examined of memory for the intention to respond in a particular way (default or special) and of memory for the color associations paired with the names. Memory for color associates was better overall with short than with long retention intervals and was better when special (rather than default) responses were required, especially at the long retention interval. These results imply that the requirement to respond in a special way protects associations from loss due to forgetting.

Two experiments examined training on a prospective time production task. Participants produced intervals, expressed in fixed arbitrary units, while performing a concurrent secondary task. After a 15-min filled delay, the participants were retrained on the same tasks. These experiments tested whether the primary and secondary tasks would be integrated into a single task. In Experiment 1, the secondary-task requirements were manipulated, but the time production task was fixed. In Experiment 2, the time production task requirements were manipulated, but the secondary task was fixed. The results suggest that participants integrate primary- and secondary-task requirements.

Two experiments examined 3 variables affecting accuracy, response time, and reports of strategy use in a binary classification skill task. In Experiment 1, higher rule cue salience, allowing faster rule application, produced higher aggregate rule use than lower rule cue salience. After participants were pretrained on the relevant classification rule, rule reports were high but generally declined across training trials; after participants were pretrained on an irrelevant rule, reports of the relevant rule increased across training trials. In Experiment 2, no rule pretraining produced a pattern of results like that obtained with irrelevant rule pretraining in Experiment 1. Presenting novel stimuli during training in Experiment 2 elevated aggregate rule reports relative to conditions where they were absent. Two participant subgroups were identified: those persisting in rule reports and those transitioning from rule to memory reports during training. The proportion of persistent rule users was higher after rule discovery than after relevant rule pretraining. Overall, the results indicate that differences among prior experiments can be reconciled. Further, they raise questions about the inevitability of memory-based automaticity in binary classification, favoring instead strategy choice based on the costs and benefits of a particular strategy and of a shift from one strategy to another.

In three experiments, we examined transfer and contextual memory in a category search task. Each experiment included two phases (training and test), during which participants searched through category and exemplar menus for targets. In Experiment 1, the targets were from one of two domains during training (grocery store or department store); the domain was either the same or changed at test. Also, the categories were organized in one of two ways (alphabetically or semantically); the organization either remained the same or changed at test. In Experiments 2 and 3, domain and organization were held constant; however, categories or exemplars were the same, partially replaced, or entirely replaced across phases in order to simulate the dynamic nature of category search in everyday situations. Transfer occurred at test when the category organization or domain was maintained and when the categories or exemplars matched (partially or entirely) those at training. These results demonstrate that transfer is facilitated by overlap in training and testing contexts.

Two experiments examined participants' responses to simulated news reports of terrorist attacks. Participants were told that a nondemocratic nation had sponsored strikes on military and cultural or educational sites in the United States. Participants in both experiments reacted more conflictually to terrorist attacks on military sites than to those on cultural or educational sites. Their conflictual responses on a thermometer scale escalated after repeated attacks. When tested in 2002 and 2004, 1 and 3 years after the real World Trade Center attacks, participants' reactions were more conflictual than those of participants examined before September 11, 2001. Furthermore, current participants' fear and anger increased, and forgiveness decreased, over repeated simulated attacks. Participants lower in masculinity showed more fear and less anger than did those higher in masculinity. This study shows that terrorist attacks produce more than simple terror.

Two experiments explored the benefits to retention and transfer conferred by mental practice. During familiarization, participants typed 4-digit numbers and took an immediate typing test on both old and new numbers. Participants then typed old 4-digit numbers, either physically or mentally, with either a different response configuration or the opposite hand from that used during familiarization. On a delayed test, participants physically typed both old and new numbers with the same response configuration and hand used during familiarization. Mental practice led to less retroactive interference and more transfer than did physical practice, supporting the hypothesis that mental practice strengthens an abstract representation that does not involve specific effectors., ((c) 2008 APA)

In 2 experiments, the efficacy of motor imagery for learning to type number sequences was examined. Adults practiced typing 4-digit numbers. Then, during subsequent training, they either typed in the same or a different location, imagined typing, merely looked at each number, or performed an irrelevant task. Repetition priming (faster responses for old relative to new numbers) was observed on an immediate test and after a 3-month delay for participants who imagined typing. Improvement across the delay in typing old and new numbers was found for the imagined and actual typing conditions but not for the other conditions. The findings suggest that imagery can be used to acquire and retain representations of sequences and to improve general typing skill., ((c) 2007 APA, all rights reserved.)

In 3 experiments, participants, on signal, moved a cursor from a central position to 1 of 8 numerically labeled locations on the circumference of a clock face. Movements were controlled by a mouse in 1 of 4 conditions: vertical reversal, horizontal reversal, combined reversals, or normal (i.e., no reversals). Participants were trained in 1, 2, or 3 of these conditions and were tested 1 week later with either the same or a different condition. There were improvements across training and perfect retention across the delay. There was little or no transfer, however, even when training involved combined reversals or multiple conditions. These results illustrate severe specificity of training and are interpreted in terms of acquired inhibition of normal responses.

Retrospective verbal protocols collected throughout participants' performance of a multiplication verification task (e.g., "7 x 3 = 28, true or false?") documented a number of different strategies and changes in strategy use across different problem categories used for this common experimental task. Correct answer retrieval and comparison to the candidate answer was the modal but not the only strategy reported. Experiment 1 results supported the use of a calculation algorithm on some trials and the use of the difference between the candidate and correct answers (i.e., split) on others. Experiment 2 clearly demonstrated that participants sometimes bypassed retrieval by relying on the split information. Implications for mental arithmetic theories and the general efficacy of retrospective protocols are discussed.

The response execution stage of cognitive skill consists of several substages, including finding the proper response location among available alternatives and moving the effector to the target location. In order to unravel the brain dynamics associated with the finding process, the present experiments used two experimental conditions. In the number condition, which requires both finding and moving, subjects are presented on each trial with a digit, 0-9, are required to find that digit on a circular clock face, and then to move a cursor to that target's location. In the arrow condition, an arrow pointing to the location of the target on the clock face circumference appears simultaneously with the target digit; no target finding is required because subjects need only to move the cursor along the path marked by the arrow. A pilot and the main experiment revealed that response initiation times but not movement times were affected by these experimental manipulations. Analysis of magnetoencephalographic (MEG) activity revealed an early occipital activity which was not affected by experimental manipulations. Later activity with central-parietal and parieto-temporal loci presumably reflected changes in the dorsal and ventral pathways, respectively, and these were affected by experimental conditions. Finally, finding processes seem to be associated with a second late activation of the ventral pathway presumably reflecting ongoing recognition processes.

We conducted two experiments to study the training of speeded responses to stimuli in a clock face array. Participants were trained on one clock face and later tested on either the same or a different clock face. The clock faces varied along 3 dimensions of change (left-right flip, up-down flip, and 180 degrees rotation). Skill acquisition was reflected in decreases in overall, initiation, and movement response times in the training and test phases except that movement time reached an asymptote during training and did not decrease further during test. Test performance was best when training and test used the same clock face and worst when the training to test change involved an up-down flip. The transition to different clock faces was consistent with an interpretation based on the violation of expectations about target positions acquired during training. Consistent trial-by-trial feedback led to better initiation time but not movement time during training than periodic feedback, but this effect did not persist into the test phase.

In 2 experiments, participants used a keyboard to enter 4-digit numbers presented on a computer monitor under conditions promoting fatigue. In Experiment 1, accuracy of data entry declined but response times improved over time, reflecting an increasing speed-accuracy trade-off. In Experiment 2, the (largely cognitive) time to enter the initial digit decreased in the 1st half but increased in the 2nd half of the session. Accuracy and time to enter the remaining digits decreased across though not within session halves. The (largely motoric) time to press a concluding keystroke decreased over the session. Thus, through a combination of facilitation and inhibition, prolonged work affects the component cognitive and motoric processes of data entry differentially and at different points in practice., ((c) 2004 APA, all rights reserved.)

N. R. Brown and R. S. Siegler (1996) found that training participants on a subset of country populations improved estimations for novel transfer country populations, an effect called seeding that remained intact over time. They attributed this effect to the abstraction by participants of a general metric framework for estimating populations not dependent on specific country anchors. In a series of 3 follow-up experiments, the authors found that training on seed populations produces both general metric information and durable specific country information. Moreover, minimal amounts of general (mean or range of populations) or specific (1 or 3 countries) information made available for inspection while estimating produced a significant seeding effect. Retention over long intervals was facilitated by both presenting 3 seed countries as opposed to 1 and providing names for the seed countries.

The training of composite skills requiring differential responding to a large set of stimuli raises issues about how to break down the whole task into parts and which parts should be trained first. Components of Morse code reception skill were identified, separated, and used to test whether initial training on a difficult part was more effective than initial training on an easy part. Initial training on a difficult subset of stimuli and on a difficult subtask both yielded disadvantages rather than the advantage implied by recent findings with different tasks. Incremental training should begin with the part yielding the most effective strategic skills, which appear to depend on characteristics of the task. In both present experiments, easy initial training led to adoption of an effective unitization strategy for representing codes. The hypothesis that procedural reinstatement at delayed testing leads to better retention was supported and extended.

It is well known that the Stroop effect is subject to influence by same-modality primes, but the possibility of cross-modal priming effects is unclear. Smell is a fundamental sensory system that is assumed to have potent cross-modality priming effects. We might expect the presence of a specific odor to interfere with performance on Stroop cards containing odor-congruent words. Forty participants, half of whom were primed with an unpleasant odor and half with a pleasant odor, were examined with modified Stroop cards containing pleasant and unpleasant descriptive words. A significant Stroop card by odor group interaction was found, indicating that the presence of an odor interferes with the performance on an odor-congruent Stroop card. These findings demonstrate cross-modality priming between olfaction and vision.

A biofeedback procedure was used to influence participants' cortical polarity before the presentation of single digit multiplication problems. To ensure that participants could solve the problems by direct memory retrieval of arithmetic facts, only nine different multiplication problems were used, and participants received extensive pre-experimental practice on these problems. After biofeedback training, cortical positivity before problem presentation was associated with faster response initiation times, a measure of mental calculation time, for correctly solved problems and with more multiplication errors than cortical negativity. Response execution time, a measure of motor speed, was not affected by cortical polarity. The shift in positivity from the amplitude of the slow cortical potential (SCP) before problem presentation to the amplitude of the positive slow wave (pSW) after problem presentation was less in positivity-required compared to negativity-required trials, mainly because of the manipulation of SCP starting points by biofeedback. We assume that cortical positivity is associated with direct retrieval of arithmetic facts, more precisely with the inhibition of incorrect answers. Cortical positivity before task presentation results in a reduced positive shift, a sign of less inhibition before answer production. The effect of less inhibition during positivity-required trials is to produce faster responses and more errors. The opposite might to be true of cortical negativity., (Copyright 1998 Elsevier Science B.V.)

Two experiments tested an identical elements model of the organization of basic arithmetic skills (T. C. Rickard, A. F. Healy, & L. E. Bourne, 1994). This model assumes a distinct abstract representation for each unique combination of the basic elements (i.e., the operands and the required operation) of a problem. Participants practiced multiplication and division problems and were then tested on various altered versions of these problems. Experiment 1 confirmed the prediction of no positive transfer when the presented elements of a test problem do not exactly match those of a practice problem. Experiment 2 confirmed the prediction that there is complete transfer when the elements of the test problem match exactly with those of a practice problem. Experiment 2 also confirmed that there is both perceptually specific and nonspecific speed-up with practice. Implications for number processing and arithmetic are discussed.

Thirteen subjects were extensively trained on nine multiplication problems varying in difficulty. Practice was associated with a reaction time speed up and an attenuation of the problem size effect. The introduction of previously unpracticed problems led to a performance rebound to pretraining levels, indicating practice specificity. The eventrelated potentials were characterized by a late positive complex, followed by a positive slow wave. Offset latency of positive slow wave and preresponse amplitude at parietal electrodes showed practice specificity effects that systematically changed with practice and problem size, indicating an association with the load imposed on working memory. The peak of the late positive complex probably reflects task learning or adaptation effects because it was attenuated by practice predominantly at frontal electrodes, showed no practice specificity, and was not affected by problem size.

Recent behavioral investigations indicate that the processes underlying mental arithmetic change systematically with practice from deliberate, conscious calculation to automatic, direct retrieval of answers from memory [Bourne, L.E.Jr. and Rickard, T.C., Mental calculation: The development of a cognitive skill, Paper presented at the Interamerican Congress of Psychology, San Jose, Costa Rica, 1991: Psychol. Rev., 95 (1988) 492-527]. Results reviewed by Moscovitch and Winocur [In: The handbook of aging and cognition, Erlbaum, Hillsdale, NJ, 1992, pp. 315-372] suggest that consciously controlled processes are more dependent on frontal lobe function than are automatic processes. It is appropriate, therefore to determine whether transitions in the locus of primary brain activity occur with practice on mental calculation. In this experiment, we examine the relationship between characteristics of event-related brain potentials (ERPs) and mental arithmetic. Single-digit mental multiplication problems varying in difficulty (problem size) were used, and subjects were trained on these problems for four sessions. Problem-size and practice effects were reliably found in behavioral measures (RT). The ERP was characterized by a pronounced late positivity after task presentation followed by a slow wave, and a negativity during response indication. These components responded differentially to the practice and problem-size manipulations. Practice mainly affected topography of the amplitude of positivity and offset latency of slow wave, and problem-size mainly offset latency of slow wave and pre-response negativity. Fronto-central positivity diminished from session to session, and the focus of positivity centered finally at centro-parietal regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Two experiments examined the relationship between implicit and explicit measures of memory for information encoded in a motoric task, called data entry. In both experiments, subjects entered lists of digit sequences with a computer keypad. They were retested on the same task after a delay of up to 1 month. At retention, implicit memory for the digit lists was evidenced by faster entry of old relative to new lists in both experiments. In Experiment 1, subjects were able to discriminate old from new lists. Recognition memory of old lists was better after than before entering the lists. In Experiment 2, perceptual and motoric contributions to the old/new difference in typing speed were isolated by means of a transfer paradigm. The results showed that the entry-speed advantage for the old lists was due to the separate reinstatement at the retention test of both perceptual and motoric procedures encoded earlier. Implicit and explicit measures of memory were found to be dependent rather than independent. The findings from this study are interpreted within a framework of memory based on procedural reinstatement.

The size of the perceptual unit used in reading was addressed using the predesignated target paradigm. Sixteen subjects viewed the following stimuli in random order: the words tee, the, tie, and toe; the nonwords eet, eht, eit, and eot; and the letters e, h, i, and o. Subjects fixated on the location of the center letter and identified the letter as e, h, i, or o, alternatives which were known to them at the onset. A word superiority effect was obtained for the common word the but not for the less common words tee, tie, and toe. The word superiority effect was attributable to bias rather than discriminability: Subjects exhibited a bias to perceive the words in this experiment as the (i.e., there was a bias to perceive h in the t e stimulus presentations). These results suggest that the common word the is processed in reading units that are larger than the letter, and that the system is biased to perceive common rather than uncommon words in data-limited conditions.

Recent evidence suggests that while pictures are more easily recognized, discriminated, associated, and recalled than their corresponding verbal labels, this is not the case in concept acquisition/utilization tasks. If such evidence is interpreted in terms of a "frequency theory" perspective, one would expect the typically obtained frequency judgment differences between pictures and words to be reduced if "conceptual" frequency judgments are required. This expectation was confirmed in three experiments.

Ss were allowed to use zero, one, two, four, or eight markers to signify the category (positive or negative) of previously selected stimuli in an attribute identification problem. The more markers available and used, the fewer trials were required for problem solution. Ss were indifferent to marker valence (positive or negative) in disjunctive problems but showed a definite preference for (the more informative) negative markers in conditionals. Performance of Ss was correlated with their tendency to move markers from lesser to more informative instances. An interpretation of results based on individual differences in information processing activities is suggested.