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\"HTTP--Hypertext transfer protocol--is the standard for exchanging messages between websites and browsers. And after 20 years, it's gotten a much-needed upgrade. With support for streams, server push, header compression, and prioritization, HTTP/2 delivers vast improvements in speed, security, and efficiency. 'HTTP/2 in action' teaches you everything you need to know to use HTTP/2 effectively. You'll learn how to optimize web performance with new features like frames, multiplexing, and push. You'll also explore real-world examples on advanced topics like flow control and dependencies. With ready-to-implement tips and best practices, this practical guide is sure to get you, and your websites, up to speed\"--Provided by publisher.

This case study investigates the innovation of redesigning the clinical trial process at a major pharmaceutical firm through the use of Lean/6-Sigma and Sprint. With the increasing complexity of protocols, there is a need to reduce the time and effort to create a Phase III protocol, the most time-consuming and costly type. This paper shows how one firm reduced from months to weeks the time to create such a protocol. Results were significant: the completed protocol was created with a 50% reduction of time with reduction of avoidable amendments and with the near elimination of process loops and email chains. Additionally, the lessons learned from this innovation were transferable to other trials.

Environmental DNA (eDNA) has seen a massive increase in application in freshwater systems with a concurrent growth in protocol developments and a drive to gain a better understanding of the ‘ecology’ of eDNA. This raises the question of whether we are currently still in an early, developmental phase of eDNA-based assessments or already transitioning into a more applied stage for biomonitoring. I conducted a systematic literature review on 381 eDNA-focused studies in freshwater systems targeting macro-organisms over the last 5 years, assessing study goals, methods, target systems and taxa and study design aspects. The results show an increase of biomonitoring-focused studies throughout the years, while the fraction of studies investigating the ‘ecology’ of eDNA decreased. The application of metabarcoding significantly increased while studies applying qPCRs tentatively declined. A geographic inequality was observed concerning study numbers and study goals biased towards the global North. Descriptive studies increased, but the fraction of in-field studies and studies applying eDNA and conventional methods combined revealed no trend. These results show a shift towards application-focused work for eDNA-based assessments but also reveal this field to still be developing. In this transitional phase, practitioners need to ensure consistency and data comparability for long-term monitoring programmes.

This paper updates previous Cochrane guidance on question formulation, searching, and protocol development, reflecting recent developments in methods for conducting qualitative evidence syntheses to inform Cochrane intervention reviews. Examples are used to illustrate how decisions about boundaries for a review are formed via an iterative process of constructing lines of inquiry and mapping the available information to ascertain whether evidence exists to answer questions related to effectiveness, implementation, feasibility, appropriateness, economic evidence, and equity. The process of question formulation allows reviewers to situate the topic in relation to how it informs and explains effectiveness, using the criterion of meaningfulness, appropriateness, feasibility, and implementation. Questions related to complex questions and interventions can be structured by drawing on an increasingly wide range of question frameworks. Logic models and theoretical frameworks are useful tools for conceptually mapping the literature to illustrate the complexity of the phenomenon of interest. Furthermore, protocol development may require iterative question formulation and searching. Consequently, the final protocol may function as a guide rather than a prescriptive route map, particularly in qualitative reviews that ask more exploratory and open-ended questions.

RNA-fluorescence in situ hybridization (FISH) is a powerful tool to visualize target messenger RNA transcripts in cultured cells, tissue sections or whole-mount preparations. As the technique has been developed over time, an ever-increasing number of divergent protocols have been published. There is now a broad selection of options available to facilitate proper tissue preparation, hybridization, and post-hybridization background removal to achieve optimal results. Here we review the technical aspects of RNA-FISH, examining the most common methods associated with different sample types including cytological preparations and whole-mounts. We discuss the application of commonly used reagents for tissue preparation, hybridization, and post-hybridization washing and provide explanations of the functional roles for each reagent. We also discuss the available probe types and necessary controls to accurately visualize gene expression. Finally, we review the most recent advances in FISH technology that facilitate both highly multiplexed experiments and signal amplification for individual targets. Taken together, this information will guide the methods development process for investigators that seek to perform FISH in organisms that lack documented or optimized protocols.

EFSA Strategy 2027 outlines the need for fit‐for‐purpose protocols for EFSA generic scientific assessments to aid in delivering trustworthy scientific advice. This EFSA Scientific Committee guidance document helps address this need by providing a harmonised and flexible framework for developing protocols for EFSA generic assessments. The guidance replaces the ‘Draft framework for protocol development for EFSA's scientific assessments’ published in 2020. The two main steps in protocol development are described. The first is problem formulation, which illustrates the objectives of the assessment. Here a new approach to translating the mandated Terms of Reference into scientifically answerable assessment questions and sub‐questions is proposed: the ‘APRIO' paradigm (Agent, Pathway, Receptor, Intervention and Output). Owing to its cross‐cutting nature, this paradigm is considered adaptable and broadly applicable within and across the various EFSA domains and, if applied using the definitions given in this guidance, is expected to help harmonise the problem formulation process and outputs and foster consistency in protocol development. APRIO may also overcome the difficulty of implementing some existing frameworks across the multiple EFSA disciplines, e.g. the PICO/PECO approach (Population, Intervention/Exposure, Comparator, Outcome). Therefore, although not mandatory, APRIO is recommended. The second step in protocol development is the specification of the evidence needs and the methods that will be applied for answering the assessment questions and sub‐questions, including uncertainty analysis. Five possible approaches to answering individual (sub‐)questions are outlined: using evidence from scientific literature and study reports; using data from databases other than bibliographic; using expert judgement informally collected or elicited via semi‐formal or formal expert knowledge elicitation processes; using mathematical/statistical models; and – not covered in this guidance – generating empirical evidence ex novo. The guidance is complemented by a standalone ‘template’ for EFSA protocols that guides the users step by step through the process of planning an EFSA scientific assessment. This publication is linked to the following EFSA Supporting Publications article: http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2022.EN-7349/full

Delays in research protocol development may be a single factor that hinders the career progression of academic faculty. Structured educational guidance during this phase proves crucial in mitigating setbacks in Institutional Review Board (IRB) approval and expediting trial implementation. To address this, the Protocol-in-a-Day (PIAD) workshop, a comprehensive 1-day event involving members from six critical facets of RO clinical trial implementation, was established, offering significant input to individual protocols. Efficacy and satisfaction of the PIAD workshop were assessed through a 5-question survey and the average time from submission to IRB initial approval. The normality of the data was analyzed using the Shapiro-Wilk Test. Nonparametric data was analyzed using a Mann-Whitney U test for significance. A total of 18 protocols that went through the PIAD workshop were activated. The mean time to IRB approval for protocols that went through PIAD was 39.8 days compared to 58.4 days for those that did not go through the PIAD workshop. Based on survey results, 100% of PIAD participants said the PIAD workshop was useful and 94% of participants stated that the PIAD workshop improved the overall quality of their protocol. Participant surveys further highlighted substantial improvements in trial quality, language, and statistical design and revealed that all participants found the workshop helpful. Therefore, both junior and senior faculty benefitted from this educational program during protocol development, as both groups demonstrated shorter times to IRB approval than non-participants. This acceleration not only fosters efficient trial implementation but also supports academic faculty in their career development.

Background A significant issue with innovative problem-solving in healthcare is an existing deficiency in continuing education for many healthcare professionals, which hinders the successful implementation of inventive solutions and progress in the field. Educators play a crucial role in guiding students to cultivate the knowledge and skills necessary to confront these challenges, including problem solving, collaboration, and the use of rapidly advancing technologies. It is vital to design educational programs that empower and motivate students to develop the proficiency and knowledge they need to be effective problem solvers, collaborators, and cultivators of innovative solutions. This project aims to assess the implementation and effectiveness of a codesigned postgraduate university program for a multidisciplinary health workforce. Methods The Leading Health Services Innovation Project is a hybrid type 2 mixed method implementation trial of a codesigned Graduate Certificate in Health Services Innovation. In collaboration with a large tertiary and quaternary health service, we developed a codesign process to guide the project, with time quarantined to create space for two-way learning between health sector partners and healthcare academics. Qualitative interviews and quantitative surveys for primary users will evaluate the implementation strategies. The reach, effectiveness, adoption implementation, and maintenance (RE-AIM) framework will guide the evaluation and maintenance of the program. Results Integrating a codesign strategy complemented by a well-structured implementation and evaluation protocol that is a combination of implementation science theoretical frameworks (Knowledge to Action, Evidence-Based Co-design, RE-AIM) may lead to translational competence as a potential outcome. Anticipated outcomes The application, resourcing and commitment to codesigned tertiary-level learning and qualification will demonstrate the achievement of a contemporary and comprehensive postgraduate university degree program in health innovation management.

•NST positively impacts the quality of care and the NS provided to critically ill surgical patients.•Education and protocol development are keystones of NST implementation.•More studies regarding NST implementation and long-term outcomes are needed for further evidence. Critically ill surgical patients pose one of the greatest challenges in achieving nutritional goals. Several published papers have demonstrated clear benefits when nutrition support (NS) is managed by a multidisciplinary nutrition support team (NST). We hypothesized that implementing a NST in a surgical intensive care unit (ICU) would increase the number of patients achieving their nutritional goals. Multicenter “BEFORE & AFTER” study. In the BEFORE phase, an audit of the previous state of NS was conducted in three ICUs without a NST. Implementation of a NST and protocol. In the AFTER phase, a new audit of NS was conducted. Continuous variables (presented as mean ± SD or median Q1–Q3) were tested using the t-test and Mann-Whitney U test. Categorical variables (presented as frequencies and percentages) were assessed using the chi-square test. A binomial logistic regression model was performed, with independent variables introduced using a stepwise forward method. A difference was considered to be significant with a two-sided P-value <0.05. Statistical analysis was conducted using IBM-SPSS 26. A total of 83 patients were included in the BEFORE phase, and 85 in the AFTER phase. The latter group showed a higher frequency of nutritional risk and malnutrition (SGA B+C odds ratio 2.314, 95% CI 1.164–4.600). Laparoscopy was more frequently utilized as a surgical technique in the AFTER phase. No differences were observed in ICU and hospital LOS or 90 days’ survival rates. Two variables remained independent factors to predict NS achievement: NST implementation (odds ratio 3.582, 95% CI 1.733–7.404), and surgical technique (odds ratio 3.231, 95% CI 1.312–7.959). NST positively impacts the chance of achieving NS goals in critically ill surgical patients.

Premise The isolation of RNA from trees is challenging due to the interference of polyphenols and polysaccharides with downstream processes. Furthermore, many RNA extraction protocols are time consuming and involve hazardous chemicals. To address these issues, we aimed to develop a safe protocol for high‐quality RNA extraction from diverse Metrosideros taxa representing a broad range of leaf toughness, pubescence, and secondary metabolites. Methods and Results We tested popular RNA isolation kits and protocols that were effective on other recalcitrant trees, including a broad range of optimization and purification steps. We optimized a protocol involving two silica‐membrane column‐based kits that yielded high‐quantity RNA with an RNA integrity number >7 and without DNA contamination. All RNA samples were used successfully in a follow‐on RNA‐Seq experiment. Conclusions We present an optimized high‐throughput RNA extraction protocol that yielded high‐quality and high‐quantity RNA from three contrasting leaf phenotypes within a hyperdiverse woody species complex.