Preparing healthcare professional students for rural, regional and remote practice: demonstrating the effectiveness of an interprofessional simulation learning experience.

Undertaking an authentic interprofessional simulation experience may be a useful and consistent strategy for healthcare professional students to build competencies required for a rural healthcare context. An observational comparative study design was adopted to evaluate a clinical simulation experience created to develop the interprofessional competencies of a sample of healthcare professional students at a regional university situated on multiple campuses in New South Wales (NSW), Australia. Over 200 students across three campuses of the university were involved in a simulation experience that included four interprofessional activities. Of these students, 189 (89%) agreed to participate in the study. The healthcare professional students who participated in the study were from second year occupational therapy, physiotherapy, and podiatry, and third year speech pathology programs. Retrospective pre and post self-assessed interprofessional collaborative competencies were compared for all students using the revised Interprofessional Collaborative Attainment Survey (ICCAS). Results demonstrated a statistically significant improvement in self-perceived scores using the validated revised ICCAS survey. The findings of this study suggest that carefully designed and authentic interprofessional simulation experiences can facilitate the development of competencies required for effective interprofessional practice, which are necessary for successful rural practice.

Preparing occupational therapy students for practice in rural areas: a scoping review protocol.

INTRODUCTION: Despite greater needs in rural areas, occupational therapists are maldistributed to urban spaces which limits service access and health outcomes for rural people and communities. Preparation of students for rural practice may improve rural workforce recruitment and retention; however, the range and scope of preparation strategies employed by education providers are unclear. This scoping review aims to understand and describe the range of strategies education services use to prepare occupational therapy students for rural practice across the globe. METHODS AND ANALYSIS: The study will include all publications about strategies used to prepare occupational therapy students for future rural practice, prior to enrolment, during the programme or on graduation. MEDLINE (Ovid), Emcare (Ovid), APA PsychInfo (Ovid), CINAHL (EBSCOhost), Health Source: Nursing/Academic (EBSCOHost), Educational Resources Information Centre (ERIC), Open Access Theses and Dissertations (OATD), and Scientific Electronic Library Online (SciELO) databases will be systematically searched in English, Spanish, French and Portuguese languages. Citations will be screened by two or more independent researchers against inclusion criteria and data extracted from included publications using a customised extraction tool. Frequency counts will be provided for study type, student location and sociodemographics, and the timing/educational strategy. The extracted data will be analysed using a matrix framework and presented in diagrammatic/tabular form and accompanied by a narrative summary which will describe how the results relate to the reviews' questions. ETHICS AND DISSEMINATION: This study will not involve human/animal subjects and does not require ethics approval. Results will be disseminated to relevant groups in peer-reviewed journal(s) and at relevant occupational therapy, higher education and/or rural health conferences. Results will also be translated and shared in blogs/social media to support access for non-research audiences and shared with other regional universities to influence curriculum design.

Profile of occupational therapy services in non-urban settings: A global scoping review.

INTRODUCTION: Regional, rural, and remote people represent nearly half the world's population yet experience disproportionally higher disease, mortality, and disability rates, coupled with limited healthcare access. Occupational therapy has committed to occupational justice, yet no descriptive framework of services provided by occupational therapists in non-urban locations exists. Understanding current non-urban service practices will provide a basis for non-urban service development and research to reduce this inequity. METHODS: Four databases were systematically searched for publications describing non-urban occupational therapy services, from any country, written in English, French, Portuguese, or Spanish, from 2010 to 2020. Publications were screened against criteria for inclusion, and data were identified using an extraction tool and presented in a frequency table, on a map, and in a searchable supporting information Table S1. RESULTS: Only 117 publications were included discussing services provided to populations across 19 countries. They were mostly published in English (98%) and about populations from English-speaking countries (70%). Included publications discussed individualist services (65%), for defined age groups (74%), and for people with specific medical diagnoses (58%). Services were commonly provided in the client's community (56%), originating from urban locations (45%) where the provider travelled (26%) or contacted clients using telehealth (19%). Individual 1:1 enabling strategies were most described (59%), including remediation (34%), compensation (9%), or education (7%). Community enablement strategies were infrequently described (14%), focused primarily on transformation to improve existing service delivery (9%), with some redistributive justice (3%), and one community development strategy (1%). Exploratory research services accounted for the remaining studies (27%). Differences were noted between Global North and South approaches. CONCLUSION: Globally, occupational therapy has limited focus on non-urban services and favours individualist rehabilitative strategies provided by therapists remote from the client's context. Further research is required on the effectiveness and appropriateness of occupational therapy strategies to improve rural/urban inequity and health outcomes.

Extension, austerity, and emergence: Themes identified from a global scoping review of non-urban occupational therapy services.

INTRODUCTION: Rural communities contribute to national wellbeing, identities, economies, and social fabrics yet experience increased risk of mortality, morbidity, and disability, coupled with lower levels of income, formal education, and employment than urban citizens. Despite higher need, occupational therapy services are maldistributed to urban locations. Publications about non-urban services discuss predominantly outreach-based, individualist, rehabilitation for specified diagnoses/age groups. However, given this population level inequity, it is unclear why individualist focussed services are more commonly discussed. Understanding intentions expressed in publications about non-urban service design may identify assumptions/limitations to current approaches and contribute to improved future services. METHODS: Each of 117 publications identified in a scoping review was read by two reviewers to independently identify themes. Provisional themes were discussed and modified in an iterative process to develop final themes/subthemes. The first author reinterrogated the publications and coded data to identify relevant examples to support the identified themes. RESULTS: Three key themes and nine subthemes were identified. Hegemonic perspectives were found in the themes (i) Extension of urban practice and (ii) Austerity, particularly in the Global North. Non-urban services were typically extended to non-urban contexts underpinned by austerity and neoliberal values such that non-urban persons and their context were problematised rather than service or funding design. A counter-hegemonic perspective was found in the theme (iii) Responses to situational realities more commonly in Global South publications, which valued non-urban contexts, and focussed on developing non-urban communities and promoting justice. CONCLUSION: The hegemonic paradigm links occupational therapy services with neoliberal notions of individualism, private provision of care, and efficiency/market value, rather than the occupational therapy values for justice. The profession must consider our role in perpetuating injustice for non-urban people and consider if and how more contextually tailored counter hegemonic place-based paradigms can be developed from and with regional, rural, and remote practice.

Identification of Natural Product Sulfuretin Derivatives as Inhibitors for the Endoplasmic Reticulum Redox Protein ERO1α.

The flavin adenine dinucleotide containing Endoplasmic Reticulum Oxidoreductase-1 α (ERO1α) catalyzes the formation of de novo disulfide bond formation of secretory and transmembrane proteins and contributes towards proper protein folding. Recently, increased ERO1α expression has been shown to contribute to increased tumor growth and metastasis in multiple cancer types. In this report we sought to define novel chemical space for targeting ERO1α function. Using the previously reported ERO1α inhibitor compound, EN-460, as a benchmark pharmacological tool we were able to identify a sulfuretin derivative, T151742 which was approximately two-fold more potent using a recombinant enzyme assay system (IC50 = 8.27 ± 2.33 µM) compared to EN-460 (IC50= 16.46 ± 3.47 µM). Additionally, T151742 (IC50 = 16.04 µM) was slightly more sensitive than EN-460 (IC50= 19.35µM) using an MTT assay as an endpoint. Utilizing a cellular thermal shift assay (CETSA), we determined that the sulfuretin derivative T151742 demonstrated isozyme specificity for ERO1α as compared to ERO1ß and showed no detectable binding to the FAD containing enzyme LSD-1. T151742 retained activity in PC-9 cells in a clonogenicity assay while EN-460 was devoid of activity. Furthermore, the activity of T151742 inhibition of clonogenicity was dependent on ERO1α expression as CRISPR edited PC-9 cells were resistant to treatment with T151742. In summary we identified a new scaffold that shows specificity for ERO1α compared to the closely related paralog ERO1ß or the FAD containing enzyme LSD-1 that can be used as a tool compound for inhibition of ERO1α to allow for pharmacological validation of the role of ERO1α in cancer.

Knee Confidence Trajectories Over Eight Years and Factors Associated With Poor Trajectories in Individuals With or at Risk for Knee Osteoarthritis.

OBJECTIVE: To identify distinct trajectories of lack of knee confidence over an 8-year follow-up period and to examine baseline factors associated with poor trajectories in individuals with or at risk for knee osteoarthritis (OA). METHODS: The Osteoarthritis Initiative is a prospective cohort study of individuals with or at high risk for knee OA. Confidence in the knees was assessed within the Knee Injury and Osteoarthritis Outcome Score instrument querying how much the individual is troubled by lack of confidence in his/her knee(s), rated as not-at-all (score = 0), mildly (score = 1), moderately (score = 2), severely (score = 3), and extremely (score = 4) troubled, reported annually from baseline to 96 months. Lack of knee confidence was defined as a score of ≥2. We used latent class models to identify subgroups that share similar underlying knee confidence trajectories over an 8-year period and multivariable multinomial logistic regression models to examine baseline factors associated with poor trajectories. RESULTS: Among 4,515 participants (mean ± SD age 61.2 ± 9.2 years, mean ± SD BMI 28.6 ± 4.8 kg/m2 ; 2,640 [58.5%] women), 4 distinct knee confidence trajectories were identified: persistently good (65.6%); declining (9.1%); poor, improving (13.9%); and persistently poor (11.4%). Baseline predictors associated with persistently poor confidence (reference: persistently good) were younger age, male sex, higher body mass index (BMI), depressive symptoms, more advanced radiographic disease, worse knee pain, weaker knee extensors, history of knee injury and surgery, and reported hip and/or ankle pain. CONCLUSION: Findings suggest the dynamic nature of self-reported knee confidence and that addressing modifiable factors (e.g., BMI, knee strength, depressive symptoms, and lower extremity pain) may improve its long-term course.

Occupational therapy services provided to populations in non-urban areas: a scoping review protocol.

OBJECTIVE: The objective of this review is to complete a global synthesis of the evidence on occupational therapy services provided in non-urban areas. INTRODUCTION: Worldwide, non-urban populations experience higher levels of disease, mortality, and disease risk factors, yet most health services, including occupational therapy, are disproportionately located in urban areas. Research has predominantly focused on attracting therapists to non-urban practice by exploring generalized expressed needs from existing non-urban therapists, such as access to professional development, backfilling of jobs, peer and other practice supports, or enticing occupational therapy students through placement experiences. However, the types of service that non-urban occupational therapists routinely provide remains unclear. Clarifying the scope of occupational therapy services in non-urban areas will support educators and policy makers to plan curricula and services to support non-urban clinicians and communities. INCLUSION CRITERIA: This scoping review will consider qualitative and quantitative research studies and opinion pieces about occupational therapy services provided to any individuals or groups living in rural (non-urban) areas across the world. METHODS: The search will be conducted in MEDLINE (Ovid), CINAHL (EBSCOhost), Emcare, and ProQuest Nursing and Allied Health, and limited to studies published from 2010 in English, French, Portuguese, or Spanish. Titles and abstracts will be screened by two independent reviewers against predefined inclusion criteria, followed by detailed assessment and appraisal by two independent reviewers. Reasons for including or excluding studies will be recorded. Data will be extracted using a charting table and presented in tabular form with a narrative summary describing how the results relate to the review question.

A Virtual Screening Platform Identifies Chloroethylagelastatin A as a Potential Ribosomal Inhibitor.

Chloroethylagelastatin A (CEAA) is an analogue of agelastatin A (AA), a natural alkaloid derived from a marine sponge. It is under development for therapeutic use against brain tumors as it has excellent central nervous system (CNS) penetration and pre-clinical therapeutic activity against brain tumors. Recently, AA was shown to inhibit protein synthesis by binding to the ribosomal A-site. In this study, we developed a novel virtual screening platform to perform a comprehensive screening of various AA analogues showing that AA analogues with proven therapeutic activity including CEAA have significant ribosomal binding capacity whereas therapeutically inactive analogues show poor ribosomal binding and revealing structural fingerprint features essential for drug-ribosome interactions. In particular, CEAA was found to have greater ribosomal binding capacity than AA. Biological tests showed that CEAA binds the ribosome and contributes to protein synthesis inhibition. Our findings suggest that CEAA may possess ribosomal inhibitor activity and that our virtual screening platform may be a useful tool in discovery and development of novel ribosomal inhibitors.

Y Chromosome LncRNA Are Involved in Radiation Response of Male Non-Small Cell Lung Cancer Cells.

Numerous studies have implicated changes in the Y chromosome in male cancers, yet few have investigated the biological importance of Y chromosome noncoding RNA. Here we identify a group of Y chromosome-expressed long noncoding RNA (lncRNA) that are involved in male non-small cell lung cancer (NSCLC) radiation sensitivity. Radiosensitive male NSCLC cell lines demonstrated a dose-dependent induction of linc-SPRY3-2/3/4 following irradiation, which was not observed in radioresistant male NSCLC cell lines. Cytogenetics revealed the loss of chromosome Y (LOY) in the radioresistant male NSCLC cell lines. Gain- and loss-of-function experiments indicated that linc-SPRY3-2/3/4 transcripts affect cell viability and apoptosis. Computational prediction of RNA binding proteins (RBP) motifs and UV-cross-linking and immunoprecipitation (CLIP) assays identified IGF2BP3, an RBP involved in mRNA stability, as a binding partner for linc-SPRY3-2/3/4 RNA. The presence of linc-SPRY3-2/3/4 reduced the half-life of known IGF2BP3 binding mRNA, such as the antiapoptotic HMGA2 mRNA, as well as the oncogenic c-MYC mRNA. Assessment of Y chromosome in NSCLC tissue microarrays and expression of linc-SPRY3-2/3/4 in NSCLC RNA-seq and microarray data revealed a negative correlation between the loss of the Y chromosome or linc-SPRY3-2/3/4 and overall survival. Thus, linc-SPRY3-2/3/4 expression and LOY could represent an important marker of radiotherapy in NSCLC. SIGNIFICANCE: This study describes previously unknown Y chromosome-expressed lncRNA regulators of radiation response in male NSCLC and show a correlation between loss of chromosome Y and radioresistance. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/19/4046/F1.large.jpg.

Effectiveness of Platelet-Rich Plasma in the Management of Knee Osteoarthritis in a Rural Clinic.

BACKGROUND: Osteoarthritis (OA) is a degenerative disease causing decreased mobility. Use of autologous platelet-rich plasma (PRP) provides a reparative alternative in the management of OA. METHODS/PURPOSE: This study assessed effectiveness of nurse practitioner-injected PRP to manage knee OA in a rural setting. RESULTS: Twenty patients were followed for 1 month after PRP treatment. Knee function and pain levels were assessed using a knee function score (Knee Injury and Osteoarthritis Outcome Score Physical Function Short Form [KOOS-PS]). CONCLUSIONS: Overall, the 20 patients who participated in this study experienced significant improvement in knee function scores, decreased pain, and decreased use of pain medications.

Long non-coding RNA FAM83H-AS1 is regulated by human papillomavirus 16 E6 independently of p53 in cervical cancer cells.

High-risk human papillomavirus (HPV) infection is one of the first events in the process of carcinogenesis in cervical and head and neck cancers. The expression of the viral oncoproteins E6 and E7 are essential in this process by inactivating the tumor suppressor proteins p53 and Rb, respectively, in addition to their interactions with other host proteins. Non-coding RNAs, such as long non-coding RNAs (lncRNAs) have been found to be dysregulated in several cancers, suggesting an important role in tumorigenesis. In order to identify host lncRNAs affected by HPV infection, we expressed the high-risk HPV-16 E6 oncoprotein in primary human keratinocytes and measured the global lncRNA expression profile by high-throughput sequencing (RNA-seq). We found several host lncRNAs differentially expressed by E6 including GAS5, H19, and FAM83H-AS1. Interestingly, FAM83H-AS1 was found overexpressed in HPV-16 positive cervical cancer cell lines in an HPV-16 E6-dependent manner but independently of p53 regulation. Furthermore, FAM83H-AS1 was found to be regulated through the E6-p300 pathway. Knockdown of FAM83H-AS1 by siRNAs decreased cellular proliferation, migration and increased apoptosis. FAM83H-AS1 was also found to be altered in human cervical cancer tissues and high expression of this lncRNA was associated with worse overall survival, suggesting an important role in cervical carcinogenesis.

Inhibition of the FAD containing ER oxidoreductin 1 (Ero1) protein by EN-460 as a strategy for treatment of multiple myeloma.

Multiple myeloma (MM) cells demonstrate high basal endoplasmic reticulum (ER) stress and are typically exquisitely sensitive to agents such as proteasome inhibitors that activate the unfolded protein response. The flavin adenosine dinucleotide (FAD) containing endoplasmic reticulum oxidoreductin enzyme (Ero1L) catalyzes de-novo disulfide bridge formation of ER resident proteins and contributes to proper protein folding. Here we show that increased Ero1L expression is prognostic of poor outcomes for MM patients relapsing on therapy. We propose that targeting protein folding via inhibition of Ero1L may represent a novel therapeutic strategy for the treatment of MM. In this report we show that treatment of MM cells with EN-460, a known inhibitor of ERO1L, was sufficient to inhibit cell proliferation and induce apoptosis. Furthermore, we show that cell death correlated in part with induction of ER stress. We also show that EN460 inhibited the enzyme activity of Ero1L, with an IC50 of 22.13 µM, consistent with previous reports. However, EN-460 was also found to inhibit other FAD-containing enzymes including MAO-A (IC50 = 7.91 µM), MAO-B (IC50 = 30.59 µM) and LSD1 (IC50 = 4.16 µM), suggesting overlap in inhibitor activity and the potential need to develop more specific inhibitors to enable pharmacological validation of ERO1L as a target for the treatment of MM. We additionally prepared and characterized azide-tagged derivatives of EN-460 as possible functional probe compounds (e.g., for photo-affinity labeling) for future target-engagement studies and further development of structure-activity relationships.

Immunoprecipitation of Tri-methylated Capped RNA.

Cellular quiescence (also known as G0 arrest) is characterized by reduced DNA replication, increased autophagy, and increased expression of cyclin-dependent kinase p27Kip1. Quiescence is essential for wound healing, organ regeneration, and preventing neoplasia. Previous findings indicate that microRNAs (miRNAs) play an important role in regulating cellular quiescence. Our recent publication demonstrated the existence of an alternative miRNA biogenesis pathway in primary human foreskin fibroblast (HFF) cells during quiescence. Indeed, we have identified a group of pri-miRNAs (whose mature miRNAs were found induced during quiescence) modified with a 2,2,7-trimethylguanosine (TMG)-cap by the trimethylguanosine synthase 1 (TGS1) protein and transported to the cytoplasm by the Exportin-1 (XPO1) protein. We used an antibody against (TMG)-caps (which does not cross-react with the (m7G)-caps that most pri-miRNAs or mRNAs contain [Luhrmann et al., 1982]) to perform RNA immunoprecipitations from total RNA extracts of proliferating or quiescent HFFs. The novelty of this assay is the specific isolation of pri-miRNAs as well as other non-coding RNAs containing a TMG-cap modification.

Widespread brain distribution and activity following i.c.v. infusion of anti-ß-secretase (BACE1) in nonhuman primates.

BACKGROUND AND PURPOSE: The potential for therapeutic antibody treatment of neurological diseases is limited by poor penetration across the blood-brain barrier. I.c.v. delivery is a promising route to the brain; however, it is unclear how efficiently antibodies delivered i.c.v. penetrate the cerebrospinal spinal fluid (CSF)-brain barrier and distribute throughout the brain parenchyma. EXPERIMENTAL APPROACH: We evaluated the pharmacokinetics and pharmacodynamics of an inhibitory monoclonal antibody against ß-secretase 1 (anti-BACE1) following continuous infusion into the left lateral ventricle of healthy adult cynomolgus monkeys. KEY RESULTS: Animals infused with anti-BACE1 i.c.v. showed a robust and sustained reduction (~70%) of CSF amyloid-ß (Aß) peptides. Antibody distribution was near uniform across the brain parenchyma, ranging from 20 to 40 nM, resulting in a ~50% reduction of Aß in the cortical parenchyma. In contrast, animals administered anti-BACE1 i.v. showed no significant change in CSF or cortical Aß levels and had a low (~0.6 nM) antibody concentration in the brain. CONCLUSION AND IMPLICATIONS: I.c.v. administration of anti-BACE1 resulted in enhanced BACE1 target engagement and inhibition, with a corresponding dramatic reduction in CNS Aß concentrations, due to enhanced brain exposure to antibody.

An Exportin-1-dependent microRNA biogenesis pathway during human cell quiescence.

The reversible state of proliferative arrest known as "cellular quiescence" plays an important role in tissue homeostasis and stem cell biology. By analyzing the expression of miRNAs and miRNA-processing factors during quiescence in primary human fibroblasts, we identified a group of miRNAs that are induced during quiescence despite markedly reduced expression of Exportin-5, a protein required for canonical miRNA biogenesis. The biogenesis of these quiescence-induced miRNAs is independent of Exportin-5 and depends instead on Exportin-1. Moreover, these quiescence-induced primary miRNAs (pri-miRNAs) are modified with a 2,2,7-trimethylguanosine (TMG)-cap, which is known to bind Exportin-1, and knockdown of Exportin-1 or trimethylguanosine synthase 1, responsible for (TMG)-capping, inhibits their biogenesis. Surprisingly, in quiescent cells Exportin-1-dependent pri-miR-34a is present in the cytoplasm together with a small isoform of Drosha, implying the existence of a different miRNA processing pathway in these cells. Our findings suggest that during quiescence the canonical miRNA biogenesis pathway is down-regulated and specific miRNAs are generated by an alternative pathway to regulate genes involved in cellular growth arrest.

Knee Instability and Basic and Advanced Function Decline in Knee Osteoarthritis.

OBJECTIVE: Manifestations of instability in knee osteoarthritis (OA) include low overall knee confidence, low confidence that the knees will not buckle, buckling, and excessive motion during gait. Confidence and buckling may particularly influence activity choices, contributing to events leading to disability. Buckling is more likely to affect advanced than basic functional tasks. In this prospective longitudinal study, we tested the hypothesis that overall knee confidence, buckling confidence, buckling, and frontal plane motion during gait are associated with advanced 2-year function outcomes in persons with knee OA. METHODS: Persons with knee OA were queried about overall knee confidence (higher score = worse confidence), buckling confidence, and knee buckling, and underwent quantitative gait analysis to quantify varus-valgus excursion and angular velocity. Physical function was assessed using the Late-Life Function and Disability Instrument Basic and Advanced Lower Extremity Domain scores. Logistic regression was used to evaluate the relationship between baseline instability measures and baseline-to-2-year function outcome, adjusting for potential confounders. RESULTS: The sample was comprised of 212 persons (mean age 64.6 years, 76.9% women). Buckling was significantly associated with poor advanced function outcome (adjusted odds ratio [OR] 2.08, 95% confidence interval [95% CI] 1.03-4.20) but not basic function outcome. Overall knee confidence was significantly associated with advanced outcome (adjusted OR 1.65, 95% CI 1.01-2.70), while associations between buckling confidence and both outcomes approached significance. Neither varus-valgus excursion nor angular velocity during gait was associated with either outcome. CONCLUSION: Knee buckling and low knee confidence were each associated with poor 2-year advanced function outcomes. Current treatment does not address these modifiable factors; interventions to address them may improve outcome in knee OA.

Factors associated with pain experience outcome in knee osteoarthritis.

OBJECTIVE: Few strategies to improve pain outcome in knee osteoarthritis (OA) exist in part because how best to evaluate pain over the long term is unclear. Our objectives were to determine the frequency of a good pain experience outcome based on previously formulated OA pain stages and test the hypothesis that less depression and pain catastrophizing and greater self-efficacy and social support are each associated with greater likelihood of a good outcome. METHODS: Study participants, all with knee OA, reported pain stage at baseline and 2 years. Baseline assessments utilized the Geriatric Depression Scale, Pain Catastrophizing Scale, Arthritis Self-Efficacy Scale, and Medical Outcomes Study social support survey. Using pain experience stages, good outcome was defined as persistence in or movement to no pain or stage 1 (predictable pain, known trigger) at 2 years. A multivariable logistic regression model was developed to identify independent predictors of a good outcome. RESULTS: Of 212 participants, 136 (64%) had a good pain outcome and 76 (36%) a poor pain outcome. In multivariable analysis, higher self-efficacy was associated with a significantly higher likelihood of good outcome (adjusted odds ratio [OR] 1.14 [95% confidence interval (95% CI) 1.04-1.24]); higher pain catastrophizing was associated with a significantly lower likelihood of good outcome (adjusted OR 0.88 [95% CI 0.83-0.94]). CONCLUSION: This stage-based measure provides a meaningful and interpretable means to assess pain outcome in knee OA. The odds of a good 2-year outcome in knee OA were lower in persons with greater pain catastrophizing and higher in persons with greater self-efficacy. Targeting these factors may help to improve pain outcome in knee OA.

Excess body weight and four-year function outcomes: comparison of African Americans and whites in a prospective study of osteoarthritis.

OBJECTIVE: We evaluated whether African Americans in the Osteoarthritis Initiative (OAI) have a greater risk (versus whites) of poor 4-year function outcome within strata defined by sex, body mass index (BMI), and waist circumference. METHODS: Using Western Ontario and McMaster Universities Osteoarthritis Index function, 20-meter walk, and chair stand performance, poor outcome was defined as moving into a worse function group or remaining in the 2 worst groups over 4 years. Logistic regression was used to evaluate the relationship between racial group and outcome within each stratum, adjusting for age, education, and income, and then further adjusting for BMI, comorbidity, depressive symptoms, physical activity, knee pain, and osteoarthritis (OA) severity. RESULTS: In 3,695 persons with or at higher risk for knee OA, higher BMI and large waist circumference were each associated with poor outcome. Among women with high BMI and among women with large waist circumference, African Americans were at greater risk for poor outcome by every measure, adjusting for age, education, and income. From fully adjusted models, potential explanatory factors included income, comorbidity, depressive symptoms, pain, and disease severity. Findings were less consistent for men, emerging only for the 20-meter walk or chair stand outcomes, and potentially explained by age and knee pain. CONCLUSION: Among OAI women with excess body weight, African Americans are at greater risk than whites for poor 4-year outcome. Modifiable factors that may help to explain these findings in the OAI include comorbidity, depressive symptoms, and knee pain. Targeting such factors, while supporting weight loss, may help to lessen the outcome disparity between African American and white women.

Quantitative measurement of invadopodia-mediated extracellular matrix proteolysis in single and multicellular contexts.

Cellular invasion into local tissues is a process important in development and homeostasis. Malregulated invasion and subsequent cell movement is characteristic of multiple pathological processes, including inflammation, cardiovascular disease and tumor cell metastasis. Focalized proteolytic degradation of extracellular matrix (ECM) components in the epithelial or endothelial basement membrane is a critical step in initiating cellular invasion. In tumor cells, extensive in vitro analysis has determined that ECM degradation is accomplished by ventral actin-rich membrane protrusive structures termed invadopodia. Invadopodia form in close apposition to the ECM, where they moderate ECM breakdown through the action of matrix metalloproteinases (MMPs). The ability of tumor cells to form invadopodia directly correlates with the ability to invade into local stroma and associated vascular components. Visualization of invadopodia-mediated ECM degradation of cells by fluorescent microscopy using dye-labeled matrix proteins coated onto glass coverslips has emerged as the most prevalent technique for evaluating the degree of matrix proteolysis and cellular invasive potential. Here we describe a version of the standard method for generating fluorescently-labeled glass coverslips utilizing a commercially available Oregon Green-488 gelatin conjugate. This method is easily scaled to rapidly produce large numbers of coated coverslips. We show some of the common microscopic artifacts that are often encountered during this procedure and how these can be avoided. Finally, we describe standardized methods using readily available computer software to allow quantification of labeled gelatin matrix degradation mediated by individual cells and by entire cellular populations. The described procedures provide the ability to accurately and reproducibly monitor invadopodia activity, and can also serve as a platform for evaluating the efficacy of modulating protein expression or testing of anti-invasive compounds on extracellular matrix degradation in single and multicellular settings.

Knee confidence as it relates to physical function outcome in persons with or at high risk of knee osteoarthritis in the osteoarthritis initiative.

OBJECTIVE: To evaluate whether low knee confidence at baseline is associated with poor baseline-to-3-year physical function outcome in the Osteoarthritis Initiative. METHODS: Knee confidence was assessed using an item from the Knee Injury and Osteoarthritis Outcome Score instrument. Physical function was assessed using self-report measures (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC] function score and Short Form 12 physical component scale) and performance-based measures (20-meter walk and chair stand test). Poor function outcome was defined as moving into a worse function group or remaining in the 2 worst function groups between baseline and 3 years. Logistic regression was used to evaluate the relationship between baseline knee confidence level and poor baseline-to-3-year function outcome, adjusting for potential confounders. RESULTS: The sample included 3,975 men and women with or at high risk of developing osteoarthritis of the knee, of whom 37-53% had poor baseline-to-3-year function outcome. For both self-report measures, increasingly worse knee confidence was associated with a greater risk of poor function outcome, and trend tests supported a graded response (e.g., the adjusted odds ratios [95% confidence intervals] for the WOMAC function score for worsening confidence categories were 1.26 [1.07-1.49], 1.43 [1.16-1.77], and 2.05 [1.49-2.82], P for trend <0.0001). Similar associations between confidence and performance-based function outcome were observed, but statistical significance did not persist in adjusted analyses. Factors independently associated with poor function outcome for all 4 outcome measures were depressive symptoms, comorbidity, body mass index, and joint space narrowing. CONCLUSION: These findings indicate that worse knee confidence at baseline is independently associated with greater risk of poor function outcome by self-report measures, with evidence of a graded response; the relationship with performance measures is not significant in fully adjusted models.