In Vivo Cellular Reprogramming: The Next Generation.

Cellular reprogramming technology has created new opportunities in understanding human disease, drug discovery, and regenerative medicine. While a combinatorial code was initially found to reprogram somatic cells to pluripotency, a "second generation" of cellular reprogramming involves lineage-restricted transcription factors and microRNAs that directly reprogram one somatic cell to another. This technology was enabled by gene networks active during development, which induce global shifts in the epigenetic landscape driving cell fate decisions. A major utility of direct reprogramming is the potential of harnessing resident support cells within damaged organs to regenerate lost tissue by converting them into the desired cell type in situ. Here, we review the progress in direct cellular reprogramming, with a focus on the paradigm of in vivo reprogramming for regenerative medicine, while pointing to hurdles that must be overcome to translate this technology into future therapeutics.

Selection of knowledge translation strategies to implement best practices on nonpharmacological prevention of delirium in intensive care unit / Sélection de stratégies de transfert des connaissances visant à implanter les pratiques recommandées de prévention non pharmacologiques du délirium en soins intensifs.

Background: Delirium prevention in the ICU should focus on a non-pharmacological approach. However, these recommendations are not always applied by care providers. Objective: To select knowledge translation strategies to facilitate the implementation of non-pharmacological best practices to prevent delirium in the ICU. Method: A consensus study was conducted. Barriers and facilitators to the implementation of nonpharmacological methods, and knowledge translation strategies, were identified in two nominal groups. A context assessment was also carried out. Nine professionals and one patient-partner participated. Results: The barriers and facilitators on which consensus was reached were most frequently related to environmental context and resources, intention, and knowledge. The areas of organizational context with the highest levels of agreement were interpersonal relations, culture and leadership. Consequently, knowledge translation strategies were selected to facilitate practices, as well as to modify the environment and improve knowledge. Conclusion: A structured method was used during this study to guide the selection of knowledge translation strategies. The application of these strategies could potentially improve clinical practice in intensive care.

Introduction: La prévention du délirium aux soins intensifs devrait être axée sur les méthodes non pharmacologiques. Toutefois, ce type de recommandation n'est pas toujours appliqué. Objectif: Sélectionner des stratégies de transfert des connaissances afin de faciliter l'implantation des pratiques non pharmacologiques pouvant prévenir le délirium en soins intensifs. Méthode: Une étude de consensus a été réalisée autour de deux thèmes. Deux groupes nominaux ont été constitués pour identifier les barrières et les facilitateurs à l'implantation des méthodes et les stratégies de transfert des connaissances. Une évaluation du contexte a aussi été réalisée. Neuf professionnels et une patiente-partenaire ont participé. Résultats: Les barrières et les facilitateurs ayant fait l'objet d'un consensus étaient plus fréquemment reliés au contexte environnemental et aux ressources, à l'intention et aux connaissances. Les domaines du contexte organisationnel qui ont obtenu le plus haut niveau d'accord sont les relations interpersonnelles, la culture et le leadership. Conséquemment, des stratégies de transfert des connaissances pour faciliter les pratiques, modifier l'environnement et améliorer les connaissances ont été sélectionnées. Conclusion: Une méthode structurée a été utilisée afin de guider la sélection de stratégies de transfert des connaissances. L'application de ces stratégies pourrait potentiellement améliorer la pratique clinique en soins intensifs.

Quantitative events determine the differentiation and function of helper T cells.

In recent years, numerous qualitative discoveries have been made in immunology research. However, the effect of quantitative events, long recognized as the driving factors for determinism in developmental biology, that dictate the quality of the immune response elicited to an antigen in concert with microbial products still requires serious attention. Here we discuss how the often-neglected issue of quantification affects the specification, differentiation and commitment of helper T cells. As reductionist in vitro approaches have been instrumental in the elucidation of the factors determining the development of helper T cells, in this perspective we highlight the need for the standardization of protocols, also fundamental for the comparison of immune responses in mice and humans. Improving understanding of how these in vitro quantitative events translate to immune responses in vivo, which can be studied in mouse models, is of importance in obtaining information on immune responses in humans, thus empowering translational research.

Improving the trustworthiness, usefulness, and ethics of biomedical research through an innovative and comprehensive institutional initiative.

The reproducibility crisis triggered worldwide initiatives to improve rigor, reproducibility, and transparency in biomedical research. There are many examples of scientists, journals, and funding agencies adopting responsible research practices. The QUEST (Quality-Ethics-Open Science-Translation) Center offers a unique opportunity to examine the role of institutions. The Berlin Institute of Health founded QUEST to increase the likelihood that research conducted at this large academic medical center would be trustworthy, useful for scientists and society, and ethical. QUEST researchers perform "science of science" studies to understand problems with standard practices and develop targeted solutions. The staff work with institutional leadership and local scientists to incentivize and support responsible practices in research, funding, and hiring. Some activities described in this paper focus on the institution, whereas others may benefit the national and international scientific community. Our experience, approaches, and recommendations will be informative for faculty leadership, administrators, and researchers interested in improving scientific practice.

Applying the science of measurement to biology: Why bother?

Both basic and translational research are continuously evolving, but the principles that underpin research integrity remain constant. These include rational, hypothesis-driven, and adequately planned and controlled science, which is carried out openly, honestly, and ethically. An important component of this should be minimising experimental irreproducibility. Biological systems, in particular, are inherently variable due to the nature of cells and tissues, as well as the complex molecules within them. As a result, it is important to understand and identify sources of variability and to strive to minimise their influence. In many instances, the application of metrology (the science of measurement) can play an important role in ensuring good quality research, even within biological systems that aren't always amenable to many of the metrological concepts applied in other fields. Here, we introduce the basic concepts of metrology in relation to biological systems and promote the application of these principles to help avoid potentially costly mistakes in both basic and translational research. We also call on funders to encourage the uptake of metrological principles, as well as provide funding and support for later engagement with regulatory bodies.

Single-cell sequencing in translational cancer research and challenges to meet clinical diagnostic needs.

The ability to capture alterations in the genome or transcriptome by next-generation sequencing has provided critical insight into molecular changes and programs underlying cancer biology. With the rapid technological development in single-cell sequencing, it has become possible to study individual cells at the transcriptional, genetic, epigenetic, and protein level. Using single-cell analysis, an increased resolution of fundamental processes underlying cancer development is obtained, providing comprehensive insights otherwise lost by sequencing of entire (bulk) samples, in which molecular signatures of individual cells are averaged across the entire cell population. Here, we provide a concise overview on the application of single-cell analysis of different modalities within cancer research by highlighting key articles of their respective fields. We furthermore examine the potential of existing technologies to meet clinical diagnostic needs and discuss current challenges associated with this translation.

Translational Assessments of Reward Responsiveness in the Marmoset.

BACKGROUND: Anhedonia, the loss of pleasure in previously rewarding activities, is a prominent feature of major depressive disorder and often resistant to first-line antidepressant treatment. A paucity of translatable cross-species tasks to assess subdomains of anhedonia, including reward learning, presents a major obstacle to the development of effective therapeutics. One assay of reward learning characterized by orderly behavioral and pharmacological findings in both humans and rats is the probabilistic reward task. In this computerized task, subjects make discriminations across numerous trials in which correct responses to one alternative are rewarded more often (rich) than correct responses to the other (lean). Healthy control subjects reliably develop a response bias to the rich alternative. However, participants with major depressive disorder as well as rats exposed to chronic stress typically exhibit a blunted response bias. METHODS: The present studies validated a touchscreen-based probabilistic reward task for the marmoset, a small nonhuman primate with considerable translational value. First, probabilistic reinforcement contingencies were parametrically examined. Next, the effects of ketamine (1.0-10.0 mg/kg), a US Food and Drug Administration-approved rapid-acting antidepressant, and phencyclidine (0.01-0.1 mg/kg), a pharmacologically similar N-methyl-D-aspartate receptor antagonist with no known antidepressant efficacy, were evaluated. RESULTS: Increases in the asymmetry of rich:lean probabilistic contingencies produced orderly increases in response bias. Consistent with their respective clinical profiles, ketamine but not phencyclidine produced dose-related increases in response bias at doses that did not reduce task discriminability. CONCLUSIONS: Collectively, these findings confirm task and pharmacological sensitivity in the marmoset, which may be useful in developing medications to counter anhedonia across neuropsychiatric disorders.

Improving natural product research translation: From source to clinical trial.

While great interest in health effects of natural product (NP) including dietary supplements and foods persists, promising preclinical NP research is not consistently translating into actionable clinical trial (CT) outcomes. Generally considered the gold standard for assessing safety and efficacy, CTs, especially phase III CTs, are costly and require rigorous planning to optimize the value of the information obtained. More effective bridging from NP research to CT was the goal of a September, 2018 transdisciplinary workshop. Participants emphasized that replicability and likelihood of successful translation depend on rigor in experimental design, interpretation, and reporting across the continuum of NP research. Discussions spanned good practices for NP characterization and quality control; use and interpretation of models (computational through in vivo) with strong clinical predictive validity; controls for experimental artefacts, especially for in vitro interrogation of bioactivity and mechanisms of action; rigorous assessment and interpretation of prior research; transparency in all reporting; and prioritization of research questions. Natural product clinical trials prioritized based on rigorous, convergent supporting data and current public health needs are most likely to be informative and ultimately affect public health. Thoughtful, coordinated implementation of these practices should enhance the knowledge gained from future NP research.

Variability in Genome Editing Outcomes: Challenges for Research Reproducibility and Clinical Safety.

Genome editing tools have already revolutionized biomedical research and are also expected to have an important impact in the clinic. However, their extensive use in research has revealed much unpredictability, both off and on target, in the outcome of their application. We discuss the challenges associated with this unpredictability, both for research and in the clinic. For the former, an extensive validation of the model is essential. For the latter, potential unpredicted activity does not preclude the use of these tools but requires that molecular evidence to underpin the relevant risk:benefit evaluation is available. Safe and successful clinical application will also depend on the mode of delivery and the cellular context.

Establishing a reproductive biorepository for basic and translational research: experience developing the reproductive subjects registry and sample repository.

PURPOSE: To report experience designing and establishing a reproductive registry and sample biorepository and to describe initial subject characteristics and biospecimens. METHODS: Beginning in December 2017, patients presenting for reproductive care at the University of Michigan were approached for study enrollment. Following consent, subjects completed detailed reproductive and health questionnaires. A variety of reproductive specimens and tissues were collected and processed for multiple downstream applications. RESULTS: Subject enrollment began in December of 2017. There are currently 1798 subjects enrolled. Female participants report a variety of reproductive disorders. Available samples include semen, sperm, follicular fluid, granulosa cells, immature oocytes, ovarian and uterine tissue, and blood samples. CONCLUSION: We report the successful establishment of a reproductive registry and sample biorepository. Furthermore, we describe methods for collection and storage of a variety of reproductive tissue processed for multiple downstream translational applications.

Implementing best available evidence into practice for incontinence-associated dermatitis in Australia: A multisite multimethod study protocol.

AIMS: Incontinence-associated dermatitis (IAD) is an insidious and under-reported hospital-acquired complication which substantially impacts on patients' quality of life. A published international guideline and the Ghent Global IAD Categorisation Tool (GLOBIAD) outline the best available evidence for the optimal management of IAD. This study aims to implement theguideline and the GLOBIAD tool and evaluate the effect on IAD occurrences and sacral pressure injuries as well as patient, clinician and cost-effectiveness outcomes. MATERIALS AND METHODS: The study will employ a multi-method design across six hospitals in five health districts in Australia, and will be conducted in three phases (pre-implementation, implementation and post-implementation) over 19 months. Data collection will involve IAD and pressure injury prevalence audits for patient hospital admissions, focus groups with, and surveys of, clinicians, patient interviews, and collection of the cost of IAD hospital care and patient-related outcomes including quality of life. Eligible participants will be hospitalised adults over 18 years of age experiencing incontinence, and clinicians working in the study wards will be invited to participate in focus groups and surveys. CONCLUSION: The implementation of health district-wide evidence-based practices for IAD using a translational research approach that engages key stakeholders will allow the standardisation of IAD care that can potentially be applicable to a range of settings. Knowledge gained will inform future practice change in patient care and health service delivery and improve the quality of care for patients with IAD. Support at the hospital, state and national levels, coupled with a refined stakeholder-inclusive strategy, will enhance this project's success, sustainability and scalability beyond this existing project.

The emergence of animal models of chronic pain and logistical and methodological issues concerning their use.

This paper examines the development of and some logistical and methodological issues surrounding the use of animal models of chronic pain. The first section addresses the emergent move towards mechanism-based and disease-related animal models of chronic pain that has accelerated since the late 1980s following publication of Bennett and Xie's (Pain 33:87-107, 1998) paper on chronic constriction injury of the sciatic nerve and Stein et al.'s (Pharmacol Biochem Behav 31:445-451, 1988) paper on unilateral hind paw inflammation with complete Freund's adjuvant. The discussion covers vast areas of chronic pain models developed over the past 50 years, starting with the numerous neuropathic, inflammatory and central pain models, as well as the growing number of models developed to study various forms of chronic pain from chronic back pain to visceral pain. It also examines the advantages and disadvantages of tonic pain models, mechanism-based and disease-related models of chronic pain, including issues related to the novel discovery of injury- or disease-related pathophysiological processes, the expansion of testing repertoires, and the successes and failures in the translation of analgesic development from animal preclinical models to human chronic pain conditions. The second section addresses experimental design considerations in the implementation of one of the 3Rs for the use of animal models of chronic pain; that is methods employed to reduce the number of animals used. The discussion covers various issues including the advantages and disadvantages of repeated dose designs and within-group drug testing, including incremental dosing schedules, and crossover designs. It also examines concerns surrounding the stability of symptoms and measures, including varying durations of multiple symptoms and the potential development of nociceptive sensitization, as well as possible use-dependent alterations in drug sensitivity and time-dependent changes in pain processes in specific animal models.

Standards and Methodological Rigor in Pulmonary Arterial Hypertension Preclinical and Translational Research.

Despite advances in our understanding of the pathophysiology and the management of pulmonary arterial hypertension (PAH), significant therapeutic gaps remain for this devastating disease. Yet, few innovative therapies beyond the traditional pathways of endothelial dysfunction have reached clinical trial phases in PAH. Although there are inherent limitations of the currently available models of PAH, the leaky pipeline of innovative therapies relates, in part, to flawed preclinical research methodology, including lack of rigour in trial design, incomplete invasive hemodynamic assessment, and lack of careful translational studies that replicate randomized controlled trials in humans with attention to adverse effects and benefits. Rigorous methodology should include the use of prespecified eligibility criteria, sample sizes that permit valid statistical analysis, randomization, blinded assessment of standardized outcomes, and transparent reporting of results. Better design and implementation of preclinical studies can minimize inherent flaws in the models of PAH, reduce the risk of bias, and enhance external validity and our ability to distinguish truly promising therapies form many false-positive or overstated leads. Ideally, preclinical studies should use advanced imaging, study several preclinical pulmonary hypertension models, or correlate rodent and human findings and consider the fate of the right ventricle, which is the major determinant of prognosis in human PAH. Although these principles are widely endorsed, empirical evidence suggests that such rigor is often lacking in pulmonary hypertension preclinical research. The present article discusses the pitfalls in the design of preclinical pulmonary hypertension trials and discusses opportunities to create preclinical trials with improved predictive value in guiding early-phase drug development in patients with PAH, which will need support not only from researchers, peer reviewers, and editors but also from academic institutions, funding agencies, and animal ethics authorities.

Standardizing serial photography for assessing and monitoring vitiligo: A core set of international recommendations for essential clinical and technical specifications.

BACKGROUND: Clinical photography is an important component of the initial assessment and follow-up of patients with vitiligo in clinical practice and research settings. Standardization of this photographic process is essential to achieve useful, high-quality, and comparable photographs over time. OBJECTIVE: The aim is to develop an international consensus for a core set of recommendations for standardized vitiligo clinical photography. METHODS: Based an international meeting of vitiligo experts, a standard operating procedure was developed for vitiligo photography in daily practice and research settings. This protocol was subsequently reviewed by 20 vitiligo experts until agreement was reached. RESULTS: The resulting protocol includes a set of 10 and 15 photographs for clinical practice and research purposes, respectively. The photographic series are based on anatomic units included in the Vitiligo Extent Score. Furthermore, graphic representations of standardized positioning and suggestions for guidelines to standardize the process (background color, lighting, position marking, scales, materials, instruments) for both color and ultraviolet photographs are described. CONCLUSIONS: This consensus-based protocol for vitiligo photography will harmonize imaging for both clinical practice, translational research, and clinical trials. It can improve outcome assessment, foster multicenter collaboration, and promote better communication with patients regarding outcomes of treatment.

Guidelines for investigating causality of sequence variants in human disease.

The discovery of rare genetic variants is accelerating, and clear guidelines for distinguishing disease-causing sequence variants from the many potentially functional variants present in any human genome are urgently needed. Without rigorous standards we risk an acceleration of false-positive reports of causality, which would impede the translation of genomic research findings into the clinical diagnostic setting and hinder biological understanding of disease. Here we discuss the key challenges of assessing sequence variants in human disease, integrating both gene-level and variant-level support for causality. We propose guidelines for summarizing confidence in variant pathogenicity and highlight several areas that require further resource development.

Technical recommendations for clinical translation of renal MRI: a consensus project of the Cooperation in Science and Technology Action PARENCHIMA.

PURPOSE: The potential of renal MRI biomarkers has been increasingly recognised, but clinical translation requires more standardisation. The PARENCHIMA consensus project aims to develop and apply a process for generating technical recommendations on renal MRI. METHODS: A task force was formed in July 2018 focused on five methods. A draft process for attaining consensus was distributed publicly for consultation and finalised at an open meeting (Prague, October 2018). Four expert panels completed surveys between October 2018 and March 2019, discussed results and refined the surveys at a face-to-face meeting (Aarhus, March 2019) and completed a second round (May 2019). RESULTS: A seven-stage process was defined: (1) formation of expert panels; (2) definition of the context of use; (3) literature review; (4) collection and comparison of MRI protocols; (5) consensus generation by an approximate Delphi method; (6) reporting of results in vendor-neutral and vendor-specific terms; (7) ongoing review and updating. Application of the process resulted in 166 consensus statements. CONCLUSION: The process generated meaningful technical recommendations across very different MRI methods, while allowing for improvement and refinement as open issues are resolved. The results are likely to be widely supported by the renal MRI community and thereby promote more harmonisation.

Building an integrated knowledge translation (IKT) evidence base: colloquium proceedings and research direction.

BACKGROUND: Integrated knowledge translation (IKT) is a model of research co-production, whereby researchers partner with knowledge users throughout the research process and who can use the research recommendations in practice or policy. IKT approaches are used to improve the relevance and impact of research. As an emerging field, however, the evidence underpinning IKT is in active development. The Integrated Knowledge Translation Research Network represents a collaborative interdisciplinary team that aims to advance the state of IKT science. METHODS: In 2017, the Integrated Knowledge Translation Research Network issued a call to its members for concept papers to further define IKT, outline an IKT research agenda, and inform the Integrated Knowledge Translation Research Network's special meeting entitled, Integrated Knowledge Translation State of the Science Colloquium, in Ottawa, Canada (2018). At the colloquium, authors presented concept papers and discussed knowledge-gaps for a research agenda and implications for advancing the IKT field. We took detailed field notes, audio-recorded the meeting and analysed the data using qualitative content analysis. RESULTS: Twenty-four participants attended the meeting, including researchers (n = 11), trainees (n = 6) and knowledge users (n = 7). Seven overarching categories emerged from these proceedings - IKT theory, IKT methods, IKT process, promoting partnership, definitions and distinctions of key IKT terms, capacity-building, and role of funders. Within these categories, priorities identified for future IKT research included: (1) improving clarity about research co-production/IKT theories and frameworks; (2) describing the process for engaging knowledge users; and (3) identifying research co-production/IKT outcomes and methods for evaluation. CONCLUSION: The Integrated Knowledge Translation State of the Science Colloquium initiated a research agenda to advance IKT science and practice. Next steps will focus on building a theoretical and evidence base for IKT.

Animal Models to Translate Phage Therapy to Human Medicine.

Phagotherapy, the use of bacteriophages to fight bacterial infections as an alternative to antibiotic treatments, has become of increasing interest in the last years. This is mainly due to the diffusion of multi-drug resistant (MDR) bacterial infections that constitute a serious issue for public health. Phage therapy is gaining favor due to its success in agriculture and veterinary treatments and its extensive utilization for human therapeutic protocols in the Eastern world. In the last decades, some clinical trials and compassionate treatments have also been performed in the Western world, indicating that phage therapy is getting closer to its introduction in standard therapy protocols. However, several questions concerning the use of phages in human therapeutic treatments are still present and need to be addressed. In this review, we illustrate the state of art of phage therapy and examine the role of animal models to translate these treatments to humans.

Putting Policy Into Practice: School-Level Compliance With and Implementation of State Concussion Laws.

CONTEXT: Each year, approximately 2 million US children 18 years or younger sustain a concussion, a type of mild traumatic brain injury (TBI). Concussions can have detrimental effects on physical, cognitive, emotional, or sleep health. POLICY: Between 2009 and 2014, all 50 US states and Washington, District of Columbia, enacted state concussion laws aimed to increase awareness about concussion and reduce the prevalence and severity of this injury. Most state laws include the following core tenets: (1) immediate removal from play after an actual or suspected concussion; (2) medical clearance before an athlete can return to play (RTP); and (3) concussion education for athletes, parents, and coaches. IMPLEMENTATION: State concussion laws allow for substantial interpretation at the school level, resulting in considerable variation in the content of school written concussion policies and the level of implementation of state law requirements at the school level. EVALUATION: We assessed the degree of high school written concussion policy compliance with the respective state law and examined the relationship between concussion policy compliance and school-level implementation of concussion laws. Seventy-one school officials completed a semistructured telephone interview and submitted their school's written concussion policy. Of the 71 policies analyzed, most complied with the removal-from-play, RTP, and concussion education tenets (90.1%, 97.2%, and 76.1%, respectively). The majority of participants reported that their school implemented the removal-from-play (91.5%), RTP (93.0%), and concussion education (80.6%) tenets well or very well. No significant relationships were found between researcher-rated school policy compliance and school-reported implementation of state law requirements at the school level. DISCUSSION: Our findings suggest that most participating schools complied with their state concussion law and implemented law requirements well or very well. Future studies should identify facilitators and barriers to the implementation of state concussion laws at the school level.

Impact of a Formal Educational Skill-Building Program Based on the ARCC Model to Enhance Evidence-Based Practice Competency in Nurse Teams.

BACKGROUND: Implementation of evidence-based practice (EBP) is necessary for healthcare systems to improve quality, safety, patient outcomes, and costs. Yet, EBP competency is lacking in many nurses and clinicians across the country. AIM: The purpose of this initiative was to determine whether nursing teams (Executive Leader, Clinical/Mid-level Leader, and Direct Care Nurse) attending a 5-day EBP continuing education skill-building program (immersion) was an effective strategy to build EBP competence, practice, and culture sustainability over time. The Advancing Research and Clinical Practice Through Close Collaboration Model was used to guide this initiative. METHODS: A project team was assembled, including leaders with EBP expertise from the Air Force Medical Service and The Helene Fuld Health Trust National Institute for EBP in Nursing and Healthcare at The Ohio State University. Five survey instruments were used to evaluate outcomes, including Organizational Culture and Readiness for System-Wide Implementation of Evidence-Based Practice, Evidence-Based Practice Beliefs, Evidence-Based Practice Implementation, and Evidence-Based Practice Competencies, as well as the Knowledge Assessment Questionnaire test. Nursing teams were invited to participate and complete the program with the implementation of EBP projects over the following year. RESULTS: Participants' EBP knowledge, skills, competencies, and beliefs were significantly improved and sustained over 12 months. LINKING EVIDENCE TO ACTION: A team-based EBP skill-building program was an effective strategy for building EBP competence, practice, and culture. This initiative demonstrated that the direct involvement of leadership and infrastructure to support EBP were crucial factors for building and sustaining an EBP culture.