Immunolocalization of proteoglycan 4 and hyaluronan on articular cartilage

INTRODUCTION Articular cartilage is a tissue that is designed to reduce friction and be resistant to wear [1]. Proteoglycan 4 (PRG4), a mucin-like glycoprotein, is a lubricating molecule that is found in synovial fluid (SF) [2]. SF also has hyaluronan (HA), a glycosaminoglycan that forms large aggregates with proteoglycans like aggrecan [3]. Friction tests have shown that PRG4 and HA exhibit synergistic effects while lubricating cartilage [4]. HA and PRG4 have both been singularly visualized on the surface of cartilage using immunohistochemistry (IHC). The objective of this project was to co-visualize PRG4 and HA on the surface of articular cartilage. METHODS Cartilage disks (6 mm diameter) were harvested from the femoral groove of mature bovine knees. Samples were either immediately snap frozen (fresh) in OCT embedding medium or were shaken overnight in phosphate buffered saline (PBS) at 4°C to remove SF, and then soaked in 200 μL bovine SF (bSF), or PBS. These samples were then snap frozen in OCT. Samples were sliced (5 μm) and placed on slides for IHC. Slides were washed in PBS, and then fixed with paraformaldehyde. They were washed again, and endogenous peroxidase activity was blocked with H2O2. Nonspecific binding was blocked with normal goat serum. Monoclonal mouse antibody 4D6 was used as the primary antibody for PRG4 and biotinylated-HA binding protein (HABP) was used to detect HA [5.6]. Alexa Fluor 594 goat anti-mouse antibodies and streptavidin PE were used as secondary agents to label the PRG4 and HA, respectively [6]. Slides were sealed with DAPI VectaShield. The slides were imaged with a Zeiss LSM 780 confocal microscope using the 405 nm (DAPI), 488 nm (HA) and 594 nm lasers (PRG4). RESULTS HA and PRG4 were co-localized on the surface of fresh cartilage. The binding was shown to be specific, since PRG4 and HA were not observed on samples that did not receive HABP or 4D6. After shaking and soaking in PBS, neither PRG4 or HA were observed on the surface indicating that shaking successfully removed SF from the surface. HA and PRG4 were observed on the shaken samples soaked in bSF, indicating the surface could be repleted with PRG4 and HA. Figure 1. Image of the surface of: fresh sample (a), fresh sample with no secondary agents (b), shake-PBS soaked (c), and shake-bSF soaked (d). Cells labeled blue. PRG4 red, HA green. Overlap between HA and PRG4 is orange/yellow. DISCUSSION AND CONCLUSIONS These results demonstrate that PRG4 and HA are co-localized on the surface of articular cartilage. The signal for PRG4 and HA is specific, and SF can be removed and repleted via shaking and soaking samples. Additional IHC work is required, with PRG4, HA, and PRG4+HA soaks, to determine the order of deposition of PRG4 and HA on the surface of articular cartilage. Further surface interaction assays could also contribute to the understanding of the PRG4+HA interaction. REFERENCES 1. Buckwalter & Mankin. J Bone Joint Surg. 79A: 612-32, 1997. 2. Drewniak E, et al. A&R. 64: 465–473, 2012. 3. Pearl A, et al. Clin Sports Med. 24: 1-12. 2005. 4. Schmidt T, et al. A&R. 56: 882-891, 2007. 5. Abubacker S, et al. Osteoarthritis and Cartilage. 21: 186-189, 2013. 6. de la Motte & Drazba. J Histochem Cytochem. 59:252–257, 2011

Exercise Interventions for Adults with Burn Injuries

The purpose of this critically appraised topic (CAT) is to provide the highest quality of evidence available on the implementation of exercise interventions in the early burn rehabilitation phase in adult burn victims. This portfolio contains four peer-reviewed research articles from national and international journals. The study designs include one systematic review and meta analyses, one retrospective cohort study and two randomized control trials. These articles covered three types of exercise interventions including resistance training, mobility training, and physiotherapy. Overall, the clinical bottom line of this CAT is that exercise interventions in early burn rehabilitation may be effective in improving upper extremity function, muscle strength, range of motion, quality of life, and decreasing length of stay and inflammation. Further research is needed to determine the effects of early exercise interventions in adults in the burn ICU

Social Media Mining for Toxicovigilance: Automatic Monitoring of Prescription Medication Abuse from Twitter

Introduction Prescription medication overdose is the fastest growing drug-related problem in the USA. The growing nature of this problem necessitates the implementation of improved monitoring strategies for investigating the prevalence and patterns of abuse of specific medications. Objectives Our primary aims were to assess the possibility of utilizing social media as a resource for automatic monitoring of prescription medication abuse and to devise an automatic classification technique that can identify potentially abuse-indicating user posts. Methods We collected Twitter user posts (tweets) associated with three commonly abused medications (Adderall®, oxycodone, and quetiapine). We manually annotated 6400 tweets mentioning these three medications and a control medication (metformin) that is not the subject of abuse due to its mechanism of action. We performed quantitative and qualitative analyses of the annotated data to determine whether posts on Twitter contain signals of prescription medication abuse. Finally, we designed an automatic supervised classification technique to distinguish posts containing signals of medication abuse from those that do not and assessed the utility of Twitter in investigating patterns of abuse over time. Results Our analyses show that clear signals of medication abuse can be drawn from Twitter posts and the percentage of tweets containing abuse signals are significantly higher for the three case medications (Adderall®: 23 %, quetiapine: 5.0 %, oxycodone: 12 %) than the proportion for the control medication (metformin: 0.3 %). Our automatic classification approach achieves 82 % accuracy overall (medication abuse class recall: 0.51, precision: 0.41, F measure: 0.46). To illustrate the utility of automatic classification, we show how the classification data can be used to analyze abuse patterns over time. Conclusion Our study indicates that social media can be a crucial resource for obtaining abuse-related information for medications, and that automatic approaches involving supervised classification and natural language processing hold promises for essential future monitoring and intervention tasks

The Effect of Teacher Professional Development on Implementing Engineering in Elementary Schools

Increased attention on the implementation of engineering education into elementary school classrooms aims to start preparing students early for potential engineering careers. In order to efficiently and effectively add engineering concepts to the curriculum, appropriate development and facilitation of engineering design challenges are required. Therefore, professional development programs are necessary to educate teachers about engineering and how to adequately teach it. This paper explores the effects of an engineering professional development program for practicing teachers. The program included training elementary teachers about how to implement units from Engineering is Elementary (EiE) by the Science Museum of Boston into their classes. Semi-structured focus group interviews were conducted, both prior to and following the implementation of the EiE units over an academic year. The interviews were transcribed and coded using open-coding, resulting in the development of a codebook. The codes were further analyzed until salient themes emerged that can be used to improve the training and better understand how teachers integrate engineering into their classrooms. The results show that many teachers need training to learn about engineering practices, as well as pedagogical guidance on how to incorporate engineering concepts into their lessons. However, not surprisingly, limited resources such as time, money, materials, and knowledge restrict efficient curricula implementation. We believe these findings reemphasize the need for science, technology, engineering, and mathematics professional development programs to educate K–12 teachers about engineering and will be useful to others interested in integrating engineering into K–12 curricula

The Clinical Sustainability Assessment Tool: Measuring organizational capacity to promote sustainability in healthcare

BACKGROUND: Few validated assessment tools are available to increase understanding and measure factors associated with sustainment of clinical practices, an increasingly recognized need among clinicians. We describe the development of the Clinical Sustainability Assessment Tool (CSAT), designed to assess factors that contribute to sustainable practices in clinical settings. METHODS: Sixty-four participants from clinical and research fields participated in concept mapping and were recruited to brainstorm factors that lead to sustained clinical practices. Once repeated factors were removed, participants sorted items based on similarity and rated them by importance and feasibility. Using concept mapping analyses, items were grouped into meaningful domains to develop an initial tool. We then recruited pilot sites and early adopters, for a total of 286 practicing clinicians, to pilot and evaluate the tool. Individuals were recruited from clinical settings across pediatric and adult medical and surgical subspecialties. The data were analyzed using confirmatory factor analysis (CFA) to test hypothesized subscale structure in the instrument. We used root mean square error of approximation (RMSEA) and the standardized root mean square residual (SRMR) to assess fit and thus the ability of CSAT to measure the identified domains. RESULTS: The concept mapping produced sorted statements that were edited into items that could be responded to, resulting in the creation of a tool with seven determinant domains and 47 items. The pilot and CFA testing resulted in a final CSAT instrument made up 35 items, five per domain. CFA results demonstrated very good fit of the seven domain structure of the CSAT (RMSEA = 0.049; SRMR = 0.049). Usability testing indicated the CSAT is brief, easy to use, easy to learn, and does not require extensive training. Additionally, the measure scored highly (18/20) on the Psychometric and Pragmatic Evidence Rating Scale (PAPERS). The seven final CSAT domains were engaged staff and leadership, engaged stakeholders, organizational readiness, workflow integration, implementation and training, monitoring and evaluation, and outcomes and effectiveness. CONCLUSIONS: The CSAT is a new reliable assessment tool which allows for greater practical and scientific understanding of contextual factors that enable sustainable clinical practices over time

New insecticide screening platforms indicate that Mitochondrial Complex I inhibitors are susceptible to cross-resistance by mosquito P450s that metabolise pyrethroids

Fenazaquin, pyridaben, tolfenpyrad and fenpyroximate are Complex I inhibitors offering a new mode of action for insecticidal malaria vector control. However, extended exposure to pyrethroid based products such as long-lasting insecticidal nets (LLINs) has created mosquito populations that are largely pyrethroid-resistant, often with elevated levels of P450s that can metabolise and neutralise diverse substrates. To assess cross-resistance liabilities of the Complex I inhibitors, we profiled their susceptibility to metabolism by P450s associated with pyrethroid resistance in Anopheles gambiae (CYPs 6M2, 6P3, 6P4, 6P5, 9J5, 9K1, 6Z2) and An. funestus (CYP6P9a). All compounds were highly susceptible. Transgenic An. gambiae overexpressing CYP6M2 or CYP6P3 showed reduced mortality when exposed to fenpyroximate and tolfenpyrad. Mortality from fenpyroximate was also reduced in pyrethroid-resistant strains of An. gambiae (VK7 2014 and Tiassalé 13) and An. funestus (FUMOZ-R). P450 inhibitor piperonyl butoxide (PBO) significantly enhanced the efficacy of fenpyroximate and tolfenpyrad, fully restoring mortality in fenpyroximate-exposed FUMOZ-R. Overall, results suggest that in vivo and in vitro assays are a useful guide in the development of new vector control products, and that the Complex I inhibitors tested are susceptible to metabolic cross-resistance and may lack efficacy in controlling pyrethroid resistant mosquitoes

Aduhelm, a novel anti-amyloid monoclonal antibody, for the treatment of Alzheimer\u27s Disease: A comprehensive review.

Alzheimer\u27s disease (AD) is the most common form of dementia affecting millions of individuals, including family members who often take on the role of caregivers. This debilitating disease reportedly consumes 8% of the total United States healthcare expenditure, with medical and nursing outlays accounting for an estimated $290 billion. Cholinesterase inhibitors and N-methyl-D-aspartate receptor antagonists have historically been the most widely used pharmacologic therapies for patients with AD; however, these drugs are not curative. The present investigation describes the epidemiology, pathophysiology, risk factors, presentation, and current treatment of AD followed by the role of the novel monoclonal antibody, Adulhelm, in the treatment of AD. Currently, Adulhelm is the only Food and Drug Administration (FDA) approved drug that acts to slow the progression of this disease. Adulhelm is an anti-amyloid drug that functions by selectively binding amyloid aggregates in both the oligomeric and fibrillar states. Studies show Adulhelm may help to restore neurological function in patients with AD by reducing beta-amyloid plaques and reestablishing neuronal calcium permeability. At present, there is concern the magnitude of this drug\u27s benefit may only be statistically significant, although not clinically significant. Despite skepticism, Adulhelm has proven to significantly decrease amyloid in all cortical brain regions examined. With such high stakes and potential, further research into Adulhelm\u27s clinical efficacy is warranted in the treatment of AD

A scoping review of frameworks in empirical studies and a review of dissemination frameworks

BACKGROUND: The field of dissemination and implementation (D&I) research has grown immensely in recent years. However, the field of dissemination research has not coalesced to the same degree as the field of implementation research. To advance the field of dissemination research, this review aimed to (1) identify the extent to which dissemination frameworks are used in dissemination empirical studies, (2) examine how scholars define dissemination, and (3) identify key constructs from dissemination frameworks. METHODS: To achieve aims 1 and 2, we conducted a scoping review of dissemination studies published in D&I science journals. The search strategy included manuscripts published from 1985 to 2020. Articles were included if they were empirical quantitative or mixed methods studies about the dissemination of information to a professional audience. Studies were excluded if they were systematic reviews, commentaries or conceptual papers, scale-up or scale-out studies, qualitative or case studies, or descriptions of programs. To achieve aim 1, we compiled the frameworks identified in the empirical studies. To achieve aim 2, we compiled the definitions from dissemination from frameworks identified in aim 1 and from dissemination frameworks identified in a 2021 review (Tabak RG, Am J Prev Med 43:337-350, 2012). To achieve aim 3, we compile the constructs and their definitions from the frameworks. FINDINGS: Out of 6017 studies, 89 studies were included for full-text extraction. Of these, 45 (51%) used a framework to guide the study. Across the 45 studies, 34 distinct frameworks were identified, out of which 13 (38%) defined dissemination. There is a lack of consensus on the definition of dissemination. Altogether, we identified 48 constructs, divided into 4 categories: process, determinants, strategies, and outcomes. Constructs in the frameworks are not well defined. IMPLICATION FOR D&I RESEARCH: This study provides a critical step in the dissemination research literature by offering suggestions on how to define dissemination research and by cataloging and defining dissemination constructs. Strengthening these definitions and distinctions between D&I research could enhance scientific reproducibility and advance the field of dissemination research

Modifications to student quarantine policies in K-12 schools implementing multiple COVID-19 prevention strategies restores in-person education without increasing SARS-CoV-2 transmission risk, January-March 2021

OBJECTIVE: To determine whether modified K-12 student quarantine policies that allow some students to continue in-person education during their quarantine period increase schoolwide SARS-CoV-2 transmission risk following the increase in cases in winter 2020-2021. METHODS: We conducted a prospective cohort study of COVID-19 cases and close contacts among students and staff (n = 65,621) in 103 Missouri public schools. Participants were offered free, saliva-based RT-PCR testing. The projected number of school-based transmission events among untested close contacts was extrapolated from the percentage of events detected among tested asymptomatic close contacts and summed with the number of detected events for a projected total. An adjusted Cox regression model compared hazard rates of school-based SARS-CoV-2 infections between schools with a modified versus standard quarantine policy. RESULTS: From January-March 2021, a projected 23 (1%) school-based transmission events occurred among 1,636 school close contacts. There was no difference in the adjusted hazard rates of school-based SARS-CoV-2 infections between schools with a modified versus standard quarantine policy (hazard ratio = 1.00; 95% confidence interval: 0.97-1.03). DISCUSSION: School-based SARS-CoV-2 transmission was rare in 103 K-12 schools implementing multiple COVID-19 prevention strategies. Modified student quarantine policies were not associated with increased school incidence of COVID-19. Modifications to student quarantine policies may be a useful strategy for K-12 schools to safely reduce disruptions to in-person education during times of increased COVID-19 community incidence