SAPPHIRE -establishment of small animal proton and photon image-guided radiation experiments.

Thein vivoevolution of radiotherapy necessitates innovative platforms for preclinical investigation, bridging the gap between bench research and clinical applications. Understanding the nuances of radiation response, specifically tailored to proton and photon therapies, is critical for optimizing treatment outcomes. Within this context, preclinicalin vivoexperimental setups incorporating image guidance for both photon and proton therapies are pivotal, enabling the translation of findings from small animal models to clinical settings. TheSAPPHIREproject represents a milestone in this pursuit, presenting the installation of the small animal radiation therapy integrated beamline (SmART+ IB, Precision X-Ray Inc., Madison, Connecticut, USA) designed for preclinical image-guided proton and photon therapy experiments at University Proton Therapy Dresden. Through Monte Carlo simulations, low-dose on-site cone beam computed tomography imaging and quality assurance alignment protocols, the project ensures the safe and precise application of radiation, crucial for replicating clinical scenarios in small animal models. The creation of Hounsfield lookup tables and comprehensive proton and photon beam characterizations within this system enable accurate dose calculations, allowing for targeted and controlled comparison experiments. By integrating these capabilities,SAPPHIREbridges preclinical investigations and potential clinical applications, offering a platform for translational radiobiology research and cancer therapy advancements.

Barriers and Facilitators When Implementing Electronic Patient-Reported Outcome Measures at a Municipal Cancer Care Unit: A Qualitative Study.

BACKGROUND: Few qualitative studies of barriers and facilitators when implementing electronic patient-reported outcome measure (ePROM) in municipal cancer care exist within the large body of symptom assessment research. Such data, gathered from healthcare professionals' (HCPs) perspective, are central to the development and design of sustainable interventions aiming for a systematic and patient-centered symptom assessment to patients with cancer. OBJECTIVE: The aim of this study was to identify and explore barriers and facilitators, as described by HCPs, in the implementation of the ePROM application "Eir" at a municipal cancer care unit in Norway. METHODS: The study applies a qualitative method, conducting an inductive data inquiry of semistructured individual interviews and focus groups with 14 Norwegian HCPs. Analysis was inspired by thematic analysis as described by Braun and Clarke. RESULTS: The analysis revealed 3 main themes affecting the implementation of ePROM in municipal cancer care: "achieving patient-centered care," "crucial management and training," and "technological barriers." CONCLUSION: The results from this study suggest that HCPs' motivation plays a significant role when implementing ePROM. Motivation of HCPs was strongly influenced by whether the application added value to previously used symptom assessment. Hands-on management and a multiprofessional approach enabled the implementation by facilitating adaptations, training, and resources. IMPLICATIONS FOR PRACTICE: The findings show that adapting the implementation of ePROMs to patient population could be of major importance. Early integration of ePROMs in cancer care could facilitate use throughout the disease trajectory.

Involving relatives in consultations for patients with long-term illnesses: Nurses and physicians' experiences.

BACKGROUND: Due to the major changes occurring in the demographic composition of the world's population, the number of older individuals is increasing, which puts pressure on the healthcare systems in many different countries. The involvement of volunteers and family members may become necessary to fulfil a patient's needs for follow-up treatments and long-term care in their homes. AIM: This study aimed to explore how nurses and physicians experienced and addressed ethical challenges when they dealt with relatives in what have traditionally been one-on-one consultations at two Norwegian hospital outpatient clinics. RESEARCH DESIGN AND METHODS: A total of eight nurses and two physicians from two different hospitals participated in individual in-depth interviews. The transcribed interviews were analysed using an eclectic approach called 'bricolage' inspired by Kvale and Brinkmann. We combined cross-case thematic and theoretical normative analyses. ETHICAL CONSIDERATIONS: The principles of voluntariness, confidentiality, withdrawal and anonymity were respected throughout the research process. In addition, the Norwegian Social Science Data Services approved this study. FINDINGS: The findings showed that respect for a patient's autonomy was used as an argument for delimiting the relatives' access to the patient's consultations. We found that there were insufficient routines in place for inviting and involving relatives in the patient consultations in the outpatient clinics. DISCUSSION: The traditional Western attitudes towards the principles of patient autonomy will likely be challenged due to the growing need for family involvement in the care of a patient in the future. CONCLUSION: This study's description of the nurses and physicians' interpretations of 'patient autonomy' as a phenomenon uncover the need for systematic ethical deliberation in the clinical setting. On an organisational level, there is a strong indication of the need to discuss the routines that are in place to invite the next of kin to participate in such patient's healthcare consultations.

Next of kin's protracted challenges with access to relevant information and involvement opportunities.

BACKGROUND: Next of kin are considered a resource for both the patient and the health service. Need for information varies with severity and duration of health changes. A clear requirement is about what to expect upon homecoming, and what supportive services are available. The picture of relatives' access to involvement and information is still somewhat unclear. OBJECTIVE: To investigate what information, knowledge, and involvement next of kin considered important for managing their caring role and collaboration with their close relatives who experienced events that led to chronic illness. DESIGN SETTING AND METHODS: A qualitative exploratory design. Seventeen informants were recruited through various courses offered to relatives. Data were collected in 2017 from individual interviews, analyzed in an interpretative tradition, and involved qualitative content analysis. RESULTS: The results reflect a long intervening period in between the activating incident and a clarification of the situation. This period was characterized by unpreparedness for duration of anxiety and amount of energy involved in balancing the relationship. Further, the interviewees saw retrospectively that information about disease and treatment was available, but they had to find such resources themselves. Information about how to handle the situation was almost absent. Ultimately, they were disappointed over not being involved. CONCLUSION: Previously provided prospective information about the embedded anxiety in the situation and consequences for relationships, involvement in patients' services, and better communication about existing services seem to be significant. Health care professionals, especially in outpatient care, may improve their services by debating how they can implement family-oriented care in personalized treatment as usual. Focus on prospective information, early involvement, and relevant information about existing resources may empower relatives and relieve the experience of care burden.

Gene editing and clonal isolation of human induced pluripotent stem cells using CRISPR/Cas9.

Human induced pluripotent stem cells (hiPSCs) represent an ideal in vitro platform to study human genetics and biology. The recent advent of programmable nucleases makes also the human genome amenable to experimental genetics through either the correction of mutations in patient-derived iPSC lines or the de novo introduction of mutations into otherwise healthy iPSCs. The production of specific and sometimes complex genotypes in multiple cell lines requires efficient and streamlined gene editing technologies. In this article we provide protocols for gene editing in hiPSCs. We presently achieve high rates of gene editing at up to three loci using a modified iCRISPR system. This system includes a doxycycline inducible Cas9 and sgRNA/reporter plasmids for the enrichment of transfected cells by fluorescence-activated cell sorting (FACS). Here we cover the selection of target sites, vector construction, transfection, and isolation and genotyping of modified hiPSC clones.

TALEN-mediated functional correction of X-linked chronic granulomatous disease in patient-derived induced pluripotent stem cells.

X-linked chronic granulomatous disease (X-CGD) is an inherited disorder of the immune system. It is characterized by a defect in the production of reactive oxygen species (ROS) in phagocytic cells due to mutations in the NOX2 locus, which encodes gp91phox. Because the success of retroviral gene therapy for X-CGD has been hampered by insertional activation of proto-oncogenes, targeting the insertion of a gp91phox transgene into potential safe harbor sites, such as AAVS1, may represent a valid alternative. To conceptually evaluate this strategy, we generated X-CGD patient-derived induced pluripotent stem cells (iPSCs), which recapitulate the cellular disease phenotype upon granulocytic differentiation. We examined AAVS1-specific zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) for their efficacy to target the insertion of a myelo-specific gp91phox cassette to AAVS1. Probably due to their lower cytotoxicity, TALENs were more efficient than ZFNs in generating correctly targeted iPSC colonies, but all corrected iPSC clones showed no signs of mutations at the top-ten predicted off-target sites of both nucleases. Upon differentiation of the corrected X-CGD iPSCs, gp91phox mRNA levels were highly up-regulated and the derived granulocytes exhibited restored ROS production that induced neutrophil extracellular trap (NET) formation. In conclusion, we demonstrate that TALEN-mediated integration of a myelo-specific gp91phox transgene into AAVS1 of patient-derived iPSCs represents a safe and efficient way to generate autologous, functionally corrected granulocytes.

Efficient designer nuclease-based homologous recombination enables direct PCR screening for footprintless targeted human pluripotent stem cells.

Genetic engineering of human induced pluripotent stem cells (hiPSCs) via customized designer nucleases has been shown to be significantly more efficient than conventional gene targeting, but still typically depends on the introduction of additional genetic selection elements. In our study, we demonstrate the efficient nonviral and selection-independent gene targeting in human pluripotent stem cells (hPSCs). Our high efficiencies of up to 1.6% of gene-targeted hiPSCs, accompanied by a low background of randomly inserted transgenes, eliminated the need for antibiotic or fluorescence-activated cell sorting selection, and allowed the use of short donor oligonucleotides for footprintless gene editing. Gene-targeted hiPSC clones were established simply by direct PCR screening. This optimized approach allows targeted transgene integration into safe harbor sites for more predictable and robust expression and enables the straightforward generation of disease-corrected, patient-derived iPSC lines for research purposes and, ultimately, for future clinical applications.

Zinc-finger nucleases-based genome engineering to generate isogenic human cell lines.

Customized zinc-finger nucleases (ZFNs) have developed into a promising technology to precisely alter mammalian genomes for biomedical research, biotechnology, or human gene therapy. In the context of synthetic biology, the targeted integration of a transgene or reporter cassette into a "neutral site" of the human genome, such as the AAVS1 locus, permits the generation of isogenic human cell lines with two major advantages over standard genetic manipulation techniques: minimal integration site-dependent effects on the transgene and, vice versa, no functional perturbation of the host-cell transcriptome. Here we describe in detail how ZFNs can be employed to target integration of a transgene cassette into the AAVS1 locus and how to characterize the targeted cells by PCR-based genotyping.

Ethical decision-making in nursing homes: influence of organizational factors.

In this article we report findings from a qualitative study that explored how doctors and nurses in nursing homes describe professional collaboration around dying patients. The study also examined the consequences this can have for the life-prolonging treatment of patients and the care of them and their relatives. Nine doctors and 10 nurses from 10 Norwegian nursing homes were interviewed about their experience of decision-making processes on life-prolonging treatment and care. The findings reveal that the frameworks for the professional collaboration and organization of physicians and nurses prevent patient treatment and care complying with ethical considerations and the law. These conditions have a challenging impact on the care of dying patients and their relatives.

Sedation of ventilated patients in intensive care units: relatives' experiences.

AIM: This paper is a report of a study undertaken to explore the experiences of being relatives of ventilated and sedated patients in intensive care units. BACKGROUND: In the last few years, issues about medically-induced sleep as part of intensive care unit treatment have focused on the appropriate level of medication. There are no studies of the impact of relatives on assessment of sedation levels, but studies of family participation in intensive care units show that relatives see their role as more important for the patient than nurses do. METHOD: An explorative, descriptive design was adopted. In-depth interviews were conducted with eight relatives of patients in 2003. The research question was: How do relatives experience having a close family member in a state of serious illness, being treated on a ventilator, and receiving medically induced sedation and pain relief? Kvale's hermeneutical interpretation principles were used for data analysis. RESULTS: Relatives interviewed seemed to go through various phases that were specifically connected to the actual situation in the intensive care unit. The main characteristic of these phases was a pattern in which participants gradually became active caregivers for the patient. Relatives wanted to be included in discussions about treatment. CONCLUSION: Relatives want to be included as caregivers when patients are under sedation in an intensive care unit. Relatives who know the patient can help evaluate the use of sedation and pain management to assist the nurses in giving the best care and the appropriate level of sedation for a patient.