High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils

Background Serine hydrolases (SHs) are a functionally diverse family of enzymes playing pivotal roles in health and disease and have emerged as important therapeutic targets in many clinical conditions. Activity-based protein profiling (ABPP) using fluorophosphonate (FP) probes has been a powerful chemoproteomic approach in studies unveiling roles of SHs in various biological systems. ABPP utilizes cell/tissue proteomes and features the FP-warhead, linked to a fluorescent reporter for in-gel fluorescence imaging or a biotin tag for streptavidin enrichment and LC-MS/MS-based target identification. Existing ABPP approaches characterize global SH activity based on mobility in gel or MS-based target identification and cannot reveal the identity of the cell-type responsible for an individual SH activity originating from complex proteomes. Results Here, by using an activity probe with broad reactivity towards the SH family, we advance the ABPP methodology to glioma brain cryosections, enabling for the first time high-resolution confocal fluorescence imaging of global SH activity in the tumor microenvironment. Tumor-associated cell types were identified by extensive immunohistochemistry on activity probe-labeled sections. Tissue-ABPP indicated heightened SH activity in glioma vs. normal brain and unveiled activity hotspots originating from tumor-associated neutrophils (TANs), rather than tumor-associated macrophages (TAMs). Thorough optimization and validation was provided by parallel gel-based ABPP combined with LC-MS/MS-based target verification. Conclusions Our study advances the ABPP methodology to tissue sections, enabling high-resolution confocal fluorescence imaging of global SH activity in anatomically preserved complex native cellular environment. To achieve global portrait of SH activity throughout the section, a probe with broad reactivity towards the SH family members was employed. As ABPP requires no a priori knowledge of the identity of the target, we envisage no imaginable reason why the presently described approach would not work for sections regardless of species and tissue source.Peer reviewe

High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils

Background Serine hydrolases (SHs) are a functionally diverse family of enzymes playing pivotal roles in health and disease and have emerged as important therapeutic targets in many clinical conditions. Activity-based protein profiling (ABPP) using fluorophosphonate (FP) probes has been a powerful chemoproteomic approach in studies unveiling roles of SHs in various biological systems. ABPP utilizes cell/tissue proteomes and features the FP-warhead, linked to a fluorescent reporter for in-gel fluorescence imaging or a biotin tag for streptavidin enrichment and LC-MS/MS-based target identification. Existing ABPP approaches characterize global SH activity based on mobility in gel or MS-based target identification and cannot reveal the identity of the cell-type responsible for an individual SH activity originating from complex proteomes. Results Here, by using an activity probe with broad reactivity towards the SH family, we advance the ABPP methodology to glioma brain cryosections, enabling for the first time high-resolution confocal fluorescence imaging of global SH activity in the tumor microenvironment. Tumor-associated cell types were identified by extensive immunohistochemistry on activity probe-labeled sections. Tissue-ABPP indicated heightened SH activity in glioma vs. normal brain and unveiled activity hotspots originating from tumor-associated neutrophils (TANs), rather than tumor-associated macrophages (TAMs). Thorough optimization and validation was provided by parallel gel-based ABPP combined with LC-MS/MS-based target verification. Conclusions Our study advances the ABPP methodology to tissue sections, enabling high-resolution confocal fluorescence imaging of global SH activity in anatomically preserved complex native cellular environment. To achieve global portrait of SH activity throughout the section, a probe with broad reactivity towards the SH family members was employed. As ABPP requires no a priori knowledge of the identity of the target, we envisage no imaginable reason why the presently described approach would not work for sections regardless of species and tissue source.Peer reviewe

Systematic reviews of observational studies of Risk of Thrombosis and Bleeding in General and Gynecologic Surgery (ROTBIGGS) : introduction and methodology

Funding Information: The Risk of Thrombosis and Bleeding in General and Gynecologic Surgery (ROTBIGGS) project was conducted by the Clinical Urology and Epidemiology (CLUE) Working Group and supported by the Academy of Finland (309387, 340957), Sigrid Jusélius Foundation and Competitive Research Funding of the Helsinki University Hospital (TYH2019321; TYH2020248). The sponsors had no role in the analysis and interpretation of the data or the manuscript preparation, review, or approval. Funding Information: KMA received a research grant from Astra Zeneca, and is consultant for Gedeon Richter, and received reimbursement for attending a scientific meeting from GSK (Tesaro Bio). RMT received reimbursement for attending a scientific meeting from Olympus. LIL, GHG, YL, RC, ALL, VJS, IEJK, PJK, RJC, RLA, KA, KMA, IB-L, MHB, JLC, SC, PJG, HAG-P, FZG, HAG, LH, MLI-K, KMJ, PKK, NK, TPK, AJK, TK, HL, AKM, BTN, TPN, CN, SMO, SP, NP, CBBR, ARR, TS, RMT, RWMV, YW, YX, LY, JH, and KAOT have no financial conflicts of interest. GHG and RC were panel members of the European Association of Urology (EAU) ad hoc Guideline on Thromboprophylaxis in Urological Surgery. KAOT was chair of the European Association of Urology (EAU) ad hoc Guideline on Thromboprophylaxis in Urological Surgery and panel member of the American Society of Hematology (ASH) Guideline Panel on Prevention of Venous Thromboembolism (VTE) in Surgical Hospitalized Patients. Publisher Copyright: © 2021, The Author(s).Background Venous thromboembolism (VTE) and bleeding are serious and potentially fatal complications of surgical procedures. Pharmacological thromboprophylaxis decreases the risk of VTE but increases the risk of major post-operative bleeding. The decision to use pharmacologic prophylaxis therefore represents a trade-off that critically depends on the incidence of VTE and bleeding in the absence of prophylaxis. These baseline risks vary widely between procedures, but their magnitude is uncertain. Systematic reviews addressing baseline risks are scarce, needed, and require innovations in methodology. Indeed, systematic summaries of these baseline risk estimates exist neither in general nor gynecologic surgery. We will fill this knowledge gap by performing a series of systematic reviews and meta-analyses of the procedure-specific and patient risk factor stratified risk estimates in general and gynecologic surgeries. Methods We will perform comprehensive literature searches for observational studies in general and gynecologic surgery reporting symptomatic VTE or bleeding estimates. Pairs of methodologically trained reviewers will independently assess the studies for eligibility, evaluate the risk of bias by using an instrument developed for this review, and extract data. We will perform meta-analyses and modeling studies to adjust the reported risk estimates for the use of thromboprophylaxis and length of follow up. We will derive the estimates of risk from the median estimates of studies rated at the lowest risk of bias. The primary outcomes are the risk estimates of symptomatic VTE and major bleeding at 4 weeks post-operatively for each procedure stratified by patient risk factors. We will apply the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach to rate evidence certainty. Discussion This series of systematic reviews, modeling studies, and meta-analyses will inform clinicians and patients regarding the trade-off between VTE prevention and bleeding in general and gynecologic surgeries. Our work advances the standards in systematic reviews of surgical complications, including assessment of risk of bias, criteria for arriving at the best estimates of risk (including modeling of the timing of events and dealing with suboptimal data reporting), dealing with subgroups at higher and lower risk of bias, and use of the GRADE approach. Systematic review registration PROSPERO CRD42021234119Peer reviewe

Systematic reviews of observational studies of Risk of Thrombosis and Bleeding in General and Gynecologic Surgery (ROTBIGGS): introduction and methodology

Background: Venous thromboembolism (VTE) and bleeding are serious and potentially fatal complications of surgical procedures. Pharmacological thromboprophylaxis decreases the risk of VTE but increases the risk of major post-operative bleeding. The decision to use pharmacologic prophylaxis therefore represents a trade-off that critically depends on the incidence of VTE and bleeding in the absence of prophylaxis. These baseline risks vary widely between procedures, but their magnitude is uncertain. Systematic reviews addressing baseline risks are scarce, needed, and require innovations in methodology. Indeed, systematic summaries of these baseline risk estimates exist neither in general nor gynecologic surgery. We will fill this knowledge gap by performing a series of systematic reviews and meta-analyses of the procedure-specific and patient risk factor stratified risk estimates in general and gynecologic surgeries.Methods: We will perform comprehensive literature searches for observational studies in general and gynecologic surgery reporting symptomatic VTE or bleeding estimates. Pairs of methodologically trained reviewers will independently assess the studies for eligibility, evaluate the risk of bias by using an instrument developed for this review, and extract data. We will perform meta-analyses and modeling studies to adjust the reported risk estimates for the use of thromboprophylaxis and length of follow up. We will derive the estimates of risk from the median estimates of studies rated at the lowest risk of bias. The primary outcomes are the risk estimates of symptomatic VTE and major bleeding at 4 weeks post-operatively for each procedure stratified by patient risk factors. We will apply the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach to rate evidence certainty.Discussion: This series of systematic reviews, modeling studies, and meta-analyses will inform clinicians and patients regarding the trade-off between VTE prevention and bleeding in general and gynecologic surgeries. Our work advances the standards in systematic reviews of surgical complications, including assessment of risk of bias, criteria for arriving at the best estimates of risk (including modeling of the timing of events and dealing with suboptimal data reporting), dealing with subgroups at higher and lower risk of bias, and use of the GRADE approach.Systematic review registration: PROSPERO CRD42021234119</p

Teachers’ experiences of using game-based learning methods in project management higher education

Abstract The benefits and challenges of using game-based learning (GBL) methods in project management higher education have not been widely investigated. This study employs qualitative methods and builds on in-depth interview data from 22 experienced university teachers with a project management teaching background. It shows that teachers perceive beneficial consequences for students from gaming, such as increased interest and knowledge acquisition with a memorable learning experience. Teachers’ inspirations and motivations are boosted by a novel method to motivate and engage students. The perceived challenges of GBL for students manifest as increased cognitive load and stress. This paper contributes to a deeper understanding of the impact of GBL methods while identifying challenges and disadvantages, which are not as widely discussed as benefits. This research identifies experiences from project management teachers’ perspectives, covering both learners’ and instructors’ viewpoints. The findings may help teachers to decide whether GBL could be used in their courses

Cultural change in the construction industry:the role of project alliance organizational culture

Abstract Project alliances are a contemporary approach for managing complex construction projects. In order to succeed, they require establishment of trust and collaborative ways of working, calling for fundamental changes in the behaviors and values among organizational members taking part in them. The increasing use of project alliancing may therefore influence the underlying values, norms and regulations within the construction sector, and hence contribute to the cultural change within the construction industry by large. The purpose of this paper is to elaborate the understanding of the cultural change that is produced by the use of project alliances, focused on collaborative behaviors. The research particularly develops knowledge on the mechanisms through which the use of project alliances may shape the industry culture. Data were collected from the Finnish construction sector, where alliance projects are increasingly popular. Central industry representatives were interviewed, and data content analysed. The findings reveal that industry level cultural change related to project alliances is a continuously evolving process and an outcome of change events and purposeful cultural change work at multiple levels including individuals, organizations, networks and construction industry. The results therefore draw attention to the role of multi-level interactions in advancing cultural change, which has to date received limited attention. The findings on the role of individuals’ career paths and trajectories are also novel, as they implicate that individual level movement and cultural orientation may play a more significant role in cultural change dynamics within project-based industries than has been acknowledged

Organizing external stakeholder engagement in inter-organizational projects:opening the black box

Abstract External stakeholder engagement is crucial for delivering value to diverse stakeholders in inter-organizational projects, however, it is not straightforward to organize this in a way that adds value. The intra-organizational focus of previous research offers limited insights into the relevant roles, responsibilities, arrangements and activities in inter-organizational contexts. Moreover, comprehensive empirical studies of the phenomenon are rare. This study explicates how internal stakeholders organize external stakeholder engagement in inter-organizational projects. Our multiple-case study of two infrastructure projects in Northern Europe identified three organizing solutions based on governance, values and dynamism. While governance-based solutions provide an overall structure for organizing external stakeholder engagement, value-based solutions ensure genuine cooperation and dynamism-based solutions facilitate timely organizing. The study develops propositions that constitute a model of how external stakeholder engagement can be organized in inter-organizational projects. The findings have implications for project stakeholder management and mainstream stakeholder research

Institutionalization of a collaborative governance model to deliver large, inter-organizational projects

Abstract Purpose: In this study, we develop further understanding of how institutional change is created within a mature and local industry. In this pursuit, we examine how a collaborative large project governance model was institutionalized at an industrial sector-level through both industry-level activities and “institutional projects”. Design/methodology/approach: This study builds on the foundations of institutional fields and institutional change, suggesting that projects are not only shaped by their contexts but also produce institutional change themselves. We conducted extensive fieldwork on the institutionalization of a collaborative project governance model in Finland. Findings: The findings illustrate how institutional change in governance of large and complex interorganizational projects is created at the institutional field level. The institutionalized collaborative project governance model includes aspects of both relational and contractual governance. The change was facilitated by temporal links between the institutional projects as well as vertical links between the institutional projects and the field-level development programs. Originality/value: This is one of the first studies to address how a collaborative large project governance model becomes the norm at the institutional field level beyond the boundaries of an individual project or organization

Creating relational capital through socialization in project alliances

Abstract Purpose: The purpose of this paper is to elaborate the understanding of socialization in the context of temporary operations and organizational settings, using project alliance — the most contemporary approach to the management of large and complex projects — as an example. In particular, the paper also assesses how informal and formal socialization mechanisms are used to facilitate relational capital in such a setting. Design/methodology/approach: Data were collected by two case studies of complex infrastructure projects in a Northern European city. The analysis focuses on how socialization is managed across organizational interfaces within the alliance organization during the project tendering and development phase to create relational capital. Findings: The findings indicate that significant emphasis is put on socialization in project alliances. However, while in the tendering phase both informal and formal socialization mechanisms are used to create relational capital; in the development phase informal socialization mechanisms are associated with higher levels of relational capital and formal socialization mechanisms are used to maintain the level of relational capital. Originality/value: While operations and supply chain management research argues that socialization is critical to manage organizational interfaces and to create relational capital in buyer-supplier relationships, research has mainly focused on ongoing operations. This study complements the prior research by developing further insight into socialization in the context of temporary operations and organizational settings; such settings create a unique empirical context, posing different managerial challenges as the results also indicate