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The mechanistic target of rapamycin (mTOR) kinase is a component of two signaling complexes that are known as mTOR complex 1 (mTORC1) and mTORC2. We sought to identify mTOR-phosphorylated proteins that are differently expressed in clinically resected clear cell renal cell carcinoma (ccRCC) relative to pair-matched normal renal tissue. Using a proteomic array, we found N-Myc Downstream Regulated 1 (NDRG1) showed the greatest increase (3.3-fold) in phosphorylation (on Thr346) in ccRCC. This was associated with an increase in total NDRG1. RICTOR is a required subunit in mTORC2, and its knockdown decreased total and phospho-NDRG1 (Thr346) but not NDRG1 mRNA. The dual mTORC1/2 inhibitor, Torin 2, significantly reduced (by ~100%) phospho-NDRG1 (Thr346). Rapamycin is a selective mTORC1 inhibitor that had no effect on the levels of total NDRG1 or phospho-NDRG1 (Thr346). The reduction in phospho-NDRG1 (Thr346) due to the inhibition of mTORC2 corresponded with a decrease in the percentage of live cells, which was correlated with an increase in apoptosis. Rapamycin had no effect on ccRCC cell viability. Collectively, these data show that mTORC2 mediates the phosphorylation of NDRG1 (Thr346) in ccRCC. We hypothesize that RICTOR and mTORC2-mediated phosphorylation of NDRG1 (Thr346) promotes the viability of ccRCC cells.

The Canadian Institutes of Health Research - Institute of Health Services and Policy Research's (IHSPR's) Strategic Plan 2021-2026 for accelerating health system transformation is well positioned to meet the strategic priorities being outlined by many health systems in Canada and internationally (CIHR IHSPR 2021). The IHSPR Health System Impact Fellow program has been a strong influence on the embedded research and scientist program in Nova Scotia, namely, the Network of Scholars Program, which was implemented just before the pandemic. The network includes scientists and scholars from diverse academic backgrounds and skill levels including alumni of the Health System Impact Fellow program. The Network of Scholars has over 30 scholars and approximately 100 academic partners and scientists supporting embedded activities such as rapid reviews, implementation science and rapid evaluation initiatives. These embedded activities are front facing to the needs and priorities of the health system. This commentary highlights the importance of IHSPR's outlined strategic plan and direction, which are consistent with the experience and the needs for embedded supports within the Nova Scotia health system.

Engaging children and young people (CYP) with and without their parents in health research has the potential to improve the development and implementation of health interventions. However, to our knowledge, the scope of engagement activities used with this population and barriers to their engagement is unknown. The objective of this review was to identify and describe CYP engagement with and without their parents in the development and/or implementation of health interventions.This scoping review included any primary research studies reporting on engaging CYP, with or without parents, in the design and/or implementation of health interventions. Healthcare professionals had to be involved over the course of the study and the study had to take place in either community, primary or tertiary care settings. The following databases were searched in May 2017, May 2020 and June 2021: Medline (OVID), CINAHL (EBSCO) and Embase (Elsevier). Two independent reviewers screened titles, abstracts and full-text articles and used a previously piloted extraction form to extract and summarize information from the included articles.Twenty-eight articles discussing twenty-four studies were included. CYP engagement throughout the research cycle was limited. There were no observed differences in the reported presence of engagement, types of interventions or outcomes of engagement between studies engaging CYP or CYP and parents. Studies engaging CYP and parents contained limited information on how these relationships affected outcomes of engagement. Engagement was enabled primarily by the maintenance of resources and relationships among stakeholders.Although CYP engagement often influenced health intervention and implementation design, they are inconsistently engaged across the research cycle. It is unclear whether parental involvement enhances CYP engagement. Future research should consider reporting guidelines to clarify the level of CYP and/or parent engagement, and enhance CYP engagement by fostering synergistic and sustainable partnerships with key stakeholders.A parent partner with codesign experience contributed to the creation of the research questions, screened titles, abstracts and full texts, helped with data extraction and provided feedback on the manuscript.

Pneumatic micro-extrusion (PME) is a high-resolution direct-write additive manufacturing method, which has been widely utilized for the fabrication of biological tissues, structures, and organs. The PME process allows for non-contact, multi-material deposition of a wide range of functional bio-inks for tissue engineering applications. However, the PME process is inherently complex, governed by complex multi-physics phenomena. Consequently, investigation of the effects of significant process parameters and their interactions on scaffold functional properties would be inevitable. The overarching goal of this research work is to fabricate biocompatible, biodegradable, and porous bone scaffolds for the treatment of large osseous fractures. In pursuit of this goal, the objective of the work is to investigate the compressive properties of triply periodic minimal surface (TPMS) bone scaffolds, composed of polycaprolactone (PCL) and fabricated using the PME additive manufacturing process. In this study, the experimental characterization of TPMS bone scaffolds was on the basis of a designed experiment. The compressive properties of the fabricated bone scaffolds were measured using a compression testing machine. Diamond as well as Icosahedron designs led to fabricated bone scaffolds having relatively high compression moduli. The results of this study pave the way for optimal fabrication of complex bone scaffolds for the treatment of bone fractures and defects.

Pneumatic micro-extrusion (PME) is a high-resolution direct-write additive manufacturing method, which has been widely utilized for the fabrication of biological tissues, structures, and organs. The PME process allows for non-contact, multi-material deposition of a wide range of functional bio-inks for tissue engineering applications. However, the PME process is inherently complex, governed by complex multi-physics phenomena. Consequently, investigation of the effects of significant process parameters and their interactions on scaffold functional properties would be inevitable. The overarching goal of this research work is to fabricate defect-free, porous bone scaffolds for the treatment of large osseous fractures. In pursuit of this goal, the objective of the work is to forward an image-based method for the characterization of the pore size distribution as well as the dimensional properties of bone tissue scaffolds, fabricated using the PME process. The developed method will allow for detection of scaffold pores, thus quantification of pore size distribution, and ultimately assessment of the dimensional accuracy of bone tissue scaffolds. The method was validated based on specimens obtained from a single factor experiment. The specimens were composed of PCL, fabricated using the PME process. The results of this study pave the way for fabrication of complex bone scaffolds with accurate dimensional properties for the treatment of bone fractures and defects.

Pneumatic micro-extrusion (PME) is a direct-write additive manufacturing process, which has emerged as a robust, high-resolution method for the fabrication of a broad spectrum of biological tissues and organs. PME allows for noncontact multimaterial deposition of functional inks for tissue engineering applications. In spite of the advantages and engendered potential applications, the PME process is inherently complex, governed not only by complex physical phenomena but also by material–process interactions. Consequently, investigation of the influence of PME process parameters as well as the underlying physical phenomena behind material transport and deposition in PME would be inevitably a need. The overarching goal of this research work is to fabricate biocompatible, porous bone tissue scaffolds for the treatment of osseous fractures, defects, and diseases. In pursuit of this goal, the objectives of the work are: (i) to investigate the influence of seven consequential scaffold design factors and PME process parameters on the mechanical properties of fabricated bone tissue scaffolds and (ii) to explore the underlying dynamics behind material transport in the PME process, using a three-dimensional computational fluid dynamics (CFD) model. To investigate the effects of the design and process parameters, a series of experiments were designed and conducted. Layer height was identified as the most significant factor in this study. An increase in the layer height led to less overlap between subsequent layers, which allowed for more shrinkage and ultimately a reduction in scaffold diameter. In addition, print speed appeared as an influential factor in this study. An increase in the print speed resulted in a decline in linear mass density and thus in the extent of fusion between subsequent deposited layers. Besides, it was observed that there was a strong correlation between deposition mass and compression modulus. Overall, the results of this study pave the way for future investigation of PME-deposited polycaprolactone (PCL) scaffolds with optimal functional and medical properties for incorporation of stem cells toward the treatment of osseous fractures and defects.

In late January 2020, the first COVID-19 case was reported in Canada. By March 5, 2020, community spread of the virus was identified and by May 26, 2020, close to 86,000 patients had COVID-19 and 6,566 had died. As COVID-19 cases increased, provincial and territorial governments announced states of public health emergency between March 13 and 20, 2020. This paper examines Canada’s public health response to the COVID-19 pandemic during the first four months (January to May 2020) by overviewing the actions undertaken by the federal (national) and regional (provincial/territorial) governments. Canada’s jurisdictional public health structures, public health responses, technological and research endeavours, and public opinion on the pandemic measures are described. As the pandemic unravelled, the federal and provincial/territorial governments unrolled a series of stringent public health interventions and restrictions, including physical distancing and gathering size restrictions; closures of borders, schools, and non-essential businesses and services; cancellations of non-essential medical services; and limitations on visitors in hospital and long-term care facilities. In late May 2020, there was a gradual decrease in the daily numbers of new COVID-19 cases seen across most jurisdictions, which has led the provinces and territories to prepare phased re-opening. Overall, the COVID-19 pandemic in Canada and the substantial amount of formative health and policy-related data being created provide an insight on how to improve responses and better prepare for future health emergencies.

Pneumatic micro-extrusion (PME), a direct-write additive manufacturing process, has emerged as a high-resolution method for the fabrication of a broad range of biological tissues and organs. However, the PME process is intrinsically complex, governed by complex physical phenomena. Hence, investigation of the effects of consequential parameters would be an inevitable need. The goal of this research work is to fabricate biocompatible, porous bone tissue scaffolds for the treatment of osseous fractures, defects, and eventually diseases. In pursuit of this goal, the objective of this study is to investigate the influence of material deposition factors — i.e., (i) deposition head temperature, (ii) flow pressure, and (iii) infill pattern — on the mechanical performance of PME-fabricated bone scaffolds. It was observed that the deposition head temperature as well as the flow pressure significantly affected scaffold diameter (unlike scaffold height). In addition, material deposition rate increased significantly as a result of an increase in the deposition temperature; this phenomenon stems from a reduction in Polycaprolactone (PCL) viscosity. Furthermore, there was a direct correlation between the amount of deposited mass and scaffold stiffness. Overall, the results of this study pave the way for future investigation of PME-deposited PCL scaffolds with optimal functional properties for incorporation of stem cells toward the treatment of osseous fractures and defects.

Pneumatic MicroExtrusion (PME) is a direct-write additive manufacturing process, which has emerged as a robust, high-resolution method for the fabrication of a broad spectrum of biological tissues and organs. However, the PME process is intrinsically complex, governed by bio-physio-chemical phenomena as well as material-process interactions. Hence, investigation of the influence of consequential factors on bone scaffold fabrication as well as investigation of cell-scaffold interactions would be an inevitable need. The objective of the work is to investigate the biocompatibility as well as the histological properties of PME-fabricated porous bone scaffolds, composed of polycaprolactone (PCL). To achieve this objective, a media extraction of the scaffold material was tested for cytostatic or cytotoxic activity with the aim to: (i) assess the fabricated scaffolds’ feasibility of use in regenerative medicine, and (ii) determine their structural integrity in a modelled in-vivo environment. In addition, the scaffolds were inoculated with an established osteosarcoma cell line (SAOS-2) and cultured for seven days to investigate the scaffold architecture and cell integration potential. A histological examination was performed on the seeded scaffolds for further in-depth analysis of cell-scaffold interaction. Overall, the results of this study pave the way for future investigation of stem cell incorporation into PME-fabricated PCL scaffolds toward the treatment of osseous fractures and defects.

e17584 Background: Disease recurrence and progression of ovarian cancer is a common event, which is accompanied by the development of platinum-resistant or refractory disease. The presence of chemo-resistant cancer stem cells (CSCs) contributes to tumor propagation, maintenance, and treatment resistance of this difficult to treat disease. The choice of appropriate chemotherapy is critical to the management of advanced epithelial ovarian cancer. We developed a CSCs assay (ChemoID) that identifies the most effective chemotherapy for individual patients from a panel of FDA-approved chemotherapies. Methods: Ascites or interventional radiology biopsies were collected under physician order from 85 consecutive patients affected by 3rd relapsed ovarian cancer. Test results from the assay were used to treat patients with the highest cell kill drugs, taking into consideration their health status and using dose reductions, if needed. Our study assessed the correlation of the ChemoID test results to treatment outcomes independently of other biomarkers. A retrospective chart analysis and review of CT and PET scans were performed to determine patients’ outcomes for tumor response, progression-free survival (PFS), and overall survival (OS). Results: We observed that recurrent ovarian cancer patients treated with high-cell kill chemotherapy agents guided by the ChemoID drug response assay had an improvement in their median PFS and OS when compared to historical median PFS and OS and/or when compared to patients who could not receive high cell kill chemotherapies (PFS low cell kill 3.5 months vs. high cell kill 12.0 months; OS low cell kill 6.0 months vs. high cell kill 15.0 months). Conclusions: The current evidence strongly suggests that the ChemoID assay predicted the effectiveness of specific therapies, as well as improved outcomes for patients treated with assay-sensitive therapies.

Pneumatic micro-extrusion (PME) is a high-resolution direct-write additive manufacturing process, which has emerged as the process of choice for tissue engineering and biofabrication of a broad spectrum of organs and tissues (e.g., bone, aortic valve, blood vessels, human ear, and nose). Despite the advantages and host of biomedical applications engendered by the PME process — including, for example, (i) accommodation of a wide range of material viscosity (enabled via thermopneumatic material deposition), (ii) large build volume and standoff distance for tissue engineering, (iii) in situ UV curing, and (iv) high-resolution multimaterial deposition — there are intrinsically complex design, material, and process factors as well as interactions, which influence the functional properties of PME-fabricated tissues and organs. Consequently, investigation of the impact and interaction of each factor aligned with establishment of a physics-based, optimal material deposition regime is inevitably a burgeoning need. In this study, using the Taguchi design, the influence of four significant factors, i.e., layer height, infill density, infill pattern, and print speed, is investigated on the compression properties as well as the dimensional accuracy of polycaprolactone (PCL) bone scaffolds, fabricated using the PME process. Furthermore, a 3D, transient two-phase flow CFD model is forwarded with the aim to observe the flow of material within the deposition head as well as the micro-capillary (nozzle). The results of this study pave the way for further investigation of the bio-functional properties of bone scaffolds, e.g., biodegradation, cell proliferation and growth rate.

Background In the United States endometrial carcinoma is the most common female gynecologic malignancy. An average of more than 60,000 new cases of endometrial carcinomas have been diagnosed yearly over the past 5 years, with a higher incidence occurring in the central Appalachian states of Ohio and West Virginia. In the U.S., the national average of newly diagnosed endometrial carcinomas is 26.8 in every 100,000 women, while in the states of Ohio and West Virginia the average is 30.5 and 31.1 in every 100,000 women, respectively. This notable increase in the incidence of endometrial carcinomas may be due a variety of elevated risk factors including but not limited to: tobacco use, obesity, and genetic predisposition of the predominant demographic. The American Cancer Society estimates that approximately 55,000 new cases of endometrial carcinoma will be diagnosed in 2020 yet, this disease is widely considered understudied and under-represented in mainstream cancer research circles. Methods The aim of this study was to quantitate the co-expression of two DNA repair proteins poly-ADP-ribose polymerase 1 and 2 (Parp-1 and Parp-2) by enzyme- linked immuno-sorbent assay (ELISA) in 60 endometrioid endometrial tumor samples and compare their expression to matched non-malignant endometrial tissue from the same corresponding donors from central Appalachia. Results We found that Parp-1 was significantly overexpressed in endometrial carcinoma relative to corresponding normal tissue. This overexpression implicates Parp inhibition therapy as a possible treatment for the disease. Our results also found a protective effect of native Parp-2 expression in non-malignant endometrial tissue with each 1 ng/mL increase in PARP-2 concentration in normal tissue was associated with a 10 % reduction in the hazard of tumor progression (HR = 0.90; p = 0.039) and a 21 % reduction in the hazard of death (HR = 0.79; p = 0.044). Conclusions This study demonstrated the over-expression of the druggable target Parp-1 in endometrial adenocarcinoma and observed a strong negative correlation of native Parp-2 expression and disease progression via the quantification of the Parp proteins using enzyme- linked immuno-sorbent assay (ELISA) assays.

Lack of standardization of clinically compliant culture protocols of mesenchymal stem cells for re-implantation in humans have hindered clinical progress in the field of tissue regeneration to repair maxillofacial and orthopedic defects. The goal of this study was to establish a clinically relevant osteogenic protocol for collection and expansion of autologous stem cells to be used at Marshall University for re-implantation and repair of maxillofacial and orthopedic conditions. Human bone marrow (hBM) samples were collected from patients undergoing intramedullary nail fixation for closed femoral fractures. hBM mesenchymal cells were expanded by growing them first in Petri dishes for two weeks, followed by a week of culture using Perfecta 3D Hanging Drop Plates&reg, Various scaffold materials were tested and analyzed for cellular integration, vitality, and differentiation capacity of harvested hBM-MSCs including: 60/40 blend of hydroxyapatite biomatrix, Acellular bone composite discs, Allowash&reg, cancellous bone cubes, PLGA (poly lactic-co-glycolic acid), and Woven chitin derived fiber. We found that the 3D spheroid culture allowed production of hBM mesenchymal cells that retained osteoblast differentiation capacity over a monolayer culture of hBM-MSCs without the need to use chemical or hormonal modulation. We also observed that hydroxyapatite and Allowash cancellous bone scaffolds allowed better cell integration and viability properties as compared to other materials tested in this study. In conclusion, the multimodal culture methodology we developed creates actively differentiating stem-cell spheroids that can then be readily utilized in clinical practices to improve the regeneration of tissues of the head and the body.

Background: Physical activity for persons with Parkinson Disease (PD) is recommended yet little is known about the physical activity levels in this patient population. The primary aim was to assess the feasibility of using a direct measurement and self-report measure of physical activity in patients with PD. Methods: Physical activity was recorded in 11 out-patients with mild to moderate PD. An accelerometer based sensor system (SenseWear Pro Armband?) which was worn continuously over 2 days was used to measure physical activity. Minute by minute energy expenditure and steps per day were recorded. Self-report physical activity was measured using the Short QUestionnaire to ASsess Health-enhancing physical activity (SQUASH) which assessed average weekly activity. Results: Using the accelerometer based sensor system, 83% of the day was spent in sedentary activity with the majority active time spent at a light intensity (2.7 [SD 2.0] hrs/day). Self-reported mean number of hours for activities greater than 2.0 METs was 3.4 (SD 1.5) hrs/day. Although the overall time spent in activity did not differ between the accelerometer and SQUASH, partici- pants reported a higher proportion of activities at the moderate and vigorous intensities than the accelerometer recorded. Conclusions: Measurement of physical activity is a challenge in persons with PD given the disease-related symptoms. We found that, by all accounts, a self-report measure of physical activity should be complemented with a direct measure of physical activity.

Highlights • Cancer stem cells (CSCs) resist chemotherapy, thereby causing relapse of the disease. • A new chemotherapy drug response assay (ChemoID), which measures the sensitivity of CSCs to a variety of chemotherapy agents has been developed. • The ChemoID assay may assist an oncologist in making treatment decisions. • The ChemoID procedure may lower treatment costs by eliminating ineffective chemotherapies and unnecessary toxicity., Introduction Administration of ineffective anticancer therapy is associated with unnecessary toxicity and development of resistant clones. Cancer stem cells (CSCs) resist chemotherapy, thereby causing relapse of the disease. Thus, development of a test that identifies the most effective chemotherapy management offers great promise for individualized anticancer treatments. We have developed an ex vivo chemotherapy drug response assay (ChemoID®), which measures the sensitivity of CSCs as well as the bulk of tumor cells to a variety of chemotherapy agents to assist an oncologist in making treatment decisions. Methods Three patients affected by oral cancer were referred. Results Biopsy showed a well-differentiated squamous cell carcinoma (G1) in case 1, a G2 adenocarcinoma in case 2 and a G3 squamous cell carcinoma in case 3. In all of the three cases, after clinical inspection and suspicion of a diagnosis of cancer, a double biopsy was performed. One specimen was sent to the ChemoID laboratory for chemosensitivity assay and the other for histological analysis. Chemotherapy dose response curves were generated, and grouped in 3 categories: 1. No response (less than 30% cell kill), Intermediate (30–60% cell kill), and 3. Sensitive (60% cell kill or above). Conclusions This procedure may be useful in helping physicians choose an effective chemotherapy regimen for head and neck cancer patients and lower treatment costs by eliminating ineffective chemotherapies and unnecessary toxicity particularly in elderly patients.

Administration of ineffective anticancer therapy is associated with unnecessary toxicity and development of resistant clones. Cancer stem-like cells (CSLCs) resist chemotherapy, thereby causing relapse of the disease. Thus, development of a test that identifies the most effective chemotherapy management offers great promise for individualized anticancer treatments. We have developed an ex vivo chemotherapy sensitivity assay (ChemoID), which measures the sensitivity of CSLCs as well as the bulk of tumor cells to a variety of chemotherapy agents. Two patients, a 21-year old male (patient 1) and a 5-month female (patient 2), affected by anaplastic WHO grade-III ependymoma were screened using the ChemoID assay. Patient 1 was found sensitive to the combination of irinotecan and bevacizumab, which resulted in a prolonged disease progression free period of 18 months. Following recurrence, the combination of various chemotherapy drugs was tested again with the ChemoID assay. We found that benzyl isothiocyanate (BITC) greatly increased the chemosensitivity of the ependymoma cells to the combination of irinotecan and bevacizumab. After patient 1 was treated for two months with irinotecan, bevacizumab and supplements of cruciferous vegetable extracts containing BITC, we observed over 50% tumoral regression in comparison with pre-ChemoID scan as evidenced by MRI. Patient 2 was found resistant to all treatments tested and following 6 cycles of vincristine, carboplatin, cyclophosphamide, etoposide, and cisplatin in various combinations, the tumor of this patient rapidly progressed and proton beam therapy was recommended. As expected animal studies conducted with patient derived xenografts treated with ChemoID screened drugs recapitulated the clinical observation. This assay demonstrates that patients with the same histological stage and grade of cancer may vary considerably in their clinical response, suggesting that ChemoID testing which measures the sensitivity of CSLCs as well as the bulk of tumor cells to a variety of chemotherapy agents could lead to more effective and personalized anticancer treatments in the future.

e13028 Background: Glioblastoma multiforme (GBMs) are brain tumors with low survival rates due to the presence of chemoresistant Glioma Cancer Stem-Like Cells (GCSLCs). Development of a test that i...

e13012 Background: Cancer stem-like cells (CSLCs) in primary brain tumors can resist certain chemotherapies, thereby causing relapse of the disease. Thus, development of a test that identifies the ...