Human Embryonic Stem Cell-Derived Mesenchymal Stroma Cells (hES-MSCs) Engraft In Vivo and Support Hematopoiesis without Suppressing Immune Function: Implications for Off-The Shelf ES-MSC Therapies.

Mesenchymal stroma cells (MSCs) have a high potential for novel cell therapy approaches in clinical transplantation. Commonly used bone marrow-derived MSCs (BM-MSCs), however, have a restricted proliferative capacity and cultures are difficult to standardize. Recently developed human embryonic stem cell-derived mesenchymal stroma cells (hES-MSCs) might represent an alternative and unlimited source of hMSCs. We therefore compared human ES-cell-derived MSCs (hES-MP002.5 cells) to normal human bone marrow-derived MSCs (BM-MSCs). hES-MP002.5 cells had lower yet reasonable CFU-F capacity compared with BM-MSC (8±3 versus 29±13 CFU-F per 100 cells). Both cell types showed similar immunophenotypic properties, i.e. cells were positive for CD105, CD73, CD166, HLA-ABC, CD44, CD146, CD90, and negative for CD45, CD34, CD14, CD31, CD117, CD19, CD 271, SSEA-4 and HLA-DR. hES-MP002.5 cells, like BM-MSCs, could be differentiated into adipocytes, osteoblasts and chondrocytes in vitro. Neither hES-MP002.5 cells nor BM-MSCs homed to the bone marrow of immune-deficient NSG mice following intravenous transplantation, whereas intra-femoral transplantation into NSG mice resulted in engraftment for both cell types. In vitro long-term culture-initiating cell assays and in vivo co-transplantation experiments with cord blood CD34+ hematopoietic cells demonstrated furthermore that hES-MP002.5 cells, like BM-MSCs, possess potent stroma support function. In contrast to BM-MSCs, however, hES-MP002.5 cells showed no or only little activity in mixed lymphocyte cultures and phytohemagglutinin (PHA) lymphocyte stimulation assays. In summary, ES-cell derived MSCs might be an attractive unlimited source for stroma transplantation approaches without suppressing immune function

Combining Flow and Mass Cytometry in the Search for Biomarkers in Chronic Graft-versus-Host Disease

Chronic graft-versus-host disease (cGVHD) is a debilitating complication arising in around half of all patients treated with an allogeneic hematopoietic stem cell transplantation. Even though treatment of severe cGVHD has improved during recent years, it remains one of the main causes of morbidity and mortality in affected patients. Biomarkers in blood that could aid in the diagnosis and classification of cGVHD severity are needed for the development of novel treatment strategies that can alleviate symptoms and reduce the need for painful and sometimes complicated tissue biopsies. Methods that comprehensively profile complex biological systems such as the immune system can reveal unanticipated markers when used with the appropriate methods of data analysis. Here, we used mass cytometry, flow cytometry, enzyme-linked immunosorbent assay, and multiplex assays to systematically profile immune cell populations in 68 patients with varying grades of cGVHD. We identified multiple subpopulations across T, B, and NK-cell lineages that distinguished patients with cGVHD from those without cGVHD and which were associated in varying ways with severity of cGVHD. Specifically, initial flow cytometry demonstrated that patients with more severe cGVHD had lower mucosal-associated T cell frequencies, with a concomitant higher level of CD38 expression on T cells. Mass cytometry could identify unique subpopulations specific for cGVHD severity albeit with some seemingly conflicting results. For instance, patients with severe cGVHD had an increased frequency of activated B cells compared to patients with moderate cGVHD while activated B cells were found at a reduced frequency in patients with mild cGVHD compared to patients without cGVHD. Moreover, results indicate it may be possible to validate mass cytometry results with clinically viable, smaller flow cytometry panels. Finally, no differences in levels of blood soluble markers could be identified, with the exception for the semi-soluble combined marker B-cell activating factor/B cell ratio, which was increased in patients with mild cGVHD compared to patients without cGVHD. These findings suggest that interdependencies between such perturbed subpopulations of cells play a role in cGVHD pathogenesis and can serve as future diagnostic and therapeutic targets

Mesenchymal Stromal Cells Engage Complement and Complement Receptor Bearing Innate Effector Cells to Modulate Immune Responses

Infusion of human third-party mesenchymal stromal cells (MSCs) appears to be a promising therapy for acute graft-versus-host disease (aGvHD). To date, little is known about how MSCs interact with the body's innate immune system after clinical infusion. This study shows, that exposure of MSCs to blood type ABO-matched human blood activates the complement system, which triggers complement-mediated lymphoid and myeloid effector cell activation in blood. We found deposition of complement component C3-derived fragments iC3b and C3dg on MSCs and fluid-phase generation of the chemotactic anaphylatoxins C3a and C5a. MSCs bound low amounts of immunoglobulins and lacked expression of complement regulatory proteins MCP (CD46) and DAF (CD55), but were protected from complement lysis via expression of protectin (CD59). Cell-surface-opsonization and anaphylatoxin-formation triggered complement receptor 3 (CD11b/CD18)-mediated effector cell activation in blood. The complement-activating properties of individual MSCs were furthermore correlated with their potency to inhibit PBMC-proliferation in vitro, and both effector cell activation and the immunosuppressive effect could be blocked either by using complement inhibitor Compstatin or by depletion of CD14/CD11b-high myeloid effector cells from mixed lymphocyte reactions. Our study demonstrates for the first time a major role of the complement system in governing the immunomodulatory activity of MSCs and elucidates how complement activation mediates the interaction with other immune cells

Gymnasieelevers uppfattningar om det laborativa momentet i kemi

Metoder De metoder vi använt är enkätundersökning, kvalitativa intervjuer samt analys av laborationsinstruktioner med avseende på antalet frihetsgrader. Resultat Vi kom i våra resultat fram till att det finns en klar dominans av ett empiristiskt synsätt på lärande i skolans kemilaborationer. Effekten av detta har blivit att eleverna oftast bara möter en typ av arbetssätt, ett induktivt med laborationer av låga frihetsgrader. Enligt tidigare undersökningar kan detta i sin tur få till följd att eleverna kan följa givna instruktioner men inte självständigt analysera problem. Med utgångspunkt i styrdokumenten så kan gymnasieeleverna inte uppnå alla mål för högre betyg om de bara provat ett styrt arbetssätt. Våra undersökningar tyder på att eleverna inte ser någon motsättning mellan empiristisk och konstruktivistisk syn på lärande, flertalet elever är positiva till att arbeta på ett mer öppet sätt under laborationer. Med utgångspunkt från våra undersökningar och styrdokumenten för gymnasieskolan anser vi att laborationer med flera frihetsgrader måste bli vanligare i gymnasieskolan. Betydelse för läraryrket Eftersom laborationer är en stor del av kemiundervisningen så är det viktigt för eleverna att laborationerna utnyttjas i undervisningen så att elevernas läroprocess gynnas och elevernas intresse stimuleras. Det är av stor relevans för kemilärare att kritiskt granska didaktiska metoder i undervisningen så att vi kan motivera metoderna utifrån hur de bidrar till elevers lärande. Eftersom det har visat sig att det finns en klar dominans av ett arbetssätt i kemins laborativa moment är det viktigt att vi som pedagoger granskar detta arbetssätt utifrån ett lärandeperspektiv

Qualitative Observational Research in the Intensive Care Setting : A Personal Reflection on Navigating Ethical and Methodological Issues

The aim of this theoretical paper is to critically reflect on the ethical and methodological issues that arose during a study that observed nurses’ care-giving in an intensive care unit setting. The authors critically discuss the methodological and ethical issues as well as the practical realities that were encountered when evaluating a complex intervention using unstructured qualitative observations. We describe the process with negotiating access and entering into the clinical field. Moreover, we reflect on experiences related to methodological issues such as the observer role, how to construct field notes, and how to encounter ethical dilemmas and other problems when being an observer in a closed and protected setting like an intensive care unit. We argue that qualitative observations give an insider perspective when studying the conditions for health and well-being. Our experiences can be transferred to other contexts and guide researchers interested in doing qualitative observational studies.CC BY-NC 4.0Fredrika Sundberg, Research and Development Centre, Skaraborg Hospital Skövde, Lövängsvägen, Skövde 541 85, Sweden. Email: [email protected] author(s) received no financial support for the research, authorship, and/or publication of this article.</p

Spatial isolation and health during the Covid-19 pandemic : A critical discourse analysis

Highlights •People's places and spaces during isolation were polarized. •Weak socioeconomic groups were voiceless in the Swedish press. •An analysis based on the ethics of justice and care is presented. •The hegemonic discourse was constructed as the Discourse of inequity.CC BY 4.0Corresponding author: Research and Development Centre, Skaraborg Hospital Skövde, Region Västra Gotaland, Sweden. E-mail address: [email protected] (F. Sundberg).This work was supported by funding from the Nordic College of Caring Science (NCCS https://www.nccs.nu/). </p

Visitor’s Experiences of an Evidence-Based Designed Healthcare Environment in an Intensive Care Unit

Objectives: The objective of the research was to study the visitors’ experiences of different healthcare environment designs of intensive care unit (ICU) patient rooms. Background: The healthcare environment may seem frightening and overwhelming in times when life-threatening conditions affect a family member or close friend and individuals visit the patient in an ICU. A two-bed patient room was refurbished to enhance the well-being of patients and their families according to the principles of evidence-based design (EBD). No prior research has used the Person-centred ClimateQuestionnaire—Family version (PCQ-F) or the semantic environment description (SMB) in the ICUsetting. Methods: A sample of 99 visitors to critically ill patients admitted to a multidisciplinary ICU completed a questionnaire; 69 visited one of the two control rooms, while 30 visited the interventionroom. Results: For the dimension of everydayness in the PCQ-F, a significantly better experience was expressed for the intervention room (p&lt;.030); the dimension regarding the ward climate general was also perceived as higher in the intervention room (p&lt;.004). The factors of pleasantness (p&lt;.019), and complexity (p&lt;0.049), showed significant differences favoring the intervention room in the SMB, with borderline significance on the modern factor (p&lt;.061). Conclusion: Designing and implementing an enriched healthcare environment in the ICU setting increases person-centered care in relation to the patients’ visitors. This could lead to better outcomes for the visitors, for example, decreasing post-traumatic stress disorder symptoms, but this needs further investigation

Associations between healthcare environment design and adverse events in intensive care unit

Background Healthcare environment can affect health. Adverse events (AEs) are common because rapid changes in the patients' status can suddenly arise, and have serious consequences, especially in intensive care. The relationship between the design of intensive care units (ICUs) and AEs has not been fully explored. Hence, an intensive care room was refurbished with cyclic lightning, sound absorbents and unique interior, and exterior design to promote health. Aims The aim of this study was to evaluate the differences between a regular and a refurbished intensive care room in risk for AEs among critically ill patients. Design This study retrospectively evaluated associations of AEs and compared the incidence of AEs in patients who were assigned to a multidisciplinary ICU in a refurbished two‐bed patient room with patients in the control rooms between 2011 and 2018. Methods There were 1938 patients included in this study (1382 in control rooms; 556 in the intervention room). Descriptive statistics were used to present the experienced AEs. Binary logistic regressions were conducted to estimate the relationship between the intervention/control rooms and variables concerning AEs. Statistical significance was set at P &lt; 0.05. Results For the frequency of AEs, there were no significant differences between the intervention room and the control rooms (10.6% vs 11%, respectively, P &lt; 0.805). No findings indicated the intervention room (the refurbished room) had a significant influence on decreasing the number of experienced AEs in critically ill patients. Conclusions The findings revealed a low incident of AEs in both the intervention room as well as in the control rooms, lower than previously described. However, our study did not find any decreases in the AEs due to the design of the rooms. Relevance to clinical practice Further research is needed to determine the relationship between the physical environment and AEs in critically ill patients

Associations between healthcare environment design and adverse events in intensive care unit

Background: Healthcare environment can affect health. Adverse events (AEs) are common because rapid changes in the patients' status can suddenly arise, and have serious consequences, especially in intensive care. The relationship between the design of intensive care units (ICUs) and AEs has not been fully explored. Hence, an intensive care room was refurbished with cyclic lightning, sound absorbents and unique interior, and exterior design to promote health. Aims: The aim of this study was to evaluate the differences between a regular and a refurbished intensive care room in risk for AEs among critically ill patients. Design: This study retrospectively evaluated associations of AEs and compared the incidence of AEs in patients who were assigned to a multidisciplinary ICU in a refurbished two-bed patient room with patients in the control rooms between 2011 and 2018. Methods: There were 1938 patients included in this study (1382 in control rooms; 556 in the intervention room). Descriptive statistics were used to present the experi-enced AEs. Binary logistic regressions were conducted to estimate the relationship between the intervention/control rooms and variables concerning AEs. Statistical significance was set at P &lt; 0.05. Results: For the frequency of AEs, there were no significant differences between the intervention room and the control rooms (10.6% vs 11%, respectively, P &lt; 0.805). No findings indicated the intervention room (the refurbished room) had a significant influence on decreasing the number of experienced AEs in critically ill patients. Conclusions: The findings revealed a low incident of AEs in both the intervention room as well as in the control rooms, lower than previously described. However, our study did not find any decreases in the AEs due to the design of the rooms. Relevance to clinical practice: Further research is needed to determine the relation-ship between the physical environment and AEs in critically ill patients

Expansion of Gammadelta T Cells from Cord Blood: A Therapeutical Possibility

Gammadelta (γδ) T cells are found in both blood and tissues and have antiviral and antitumor properties. The frequency of γδ T cells in umbilical cord blood (UCB) is low, and the majority express δ1, in contrast to blood, whereas the main subset is δ2γ9 T cells. UCB γδ T cells are functionally immature, which together with their scarcity complicates the development of UCB γδ T cell therapies. We aimed to develop an effective expansion protocol for UCB γδ T cells based on zoledronate and IL-2. We found that culture with 5 μM zoledronate and 200 IU IL-2/ml medium for 14 days promoted extensive proliferation. The majority of the cultured cells were γ9δ2 T cells. The fold expansion of this, originally infrequent, subset was impressive (median and maximum fold change 253 and 1085, resp.). After culture, the cells had a polyclonal γδ T cell repertoire and the main memory subset was central memory (CD45RO+ CD27+). The cells produced cytokines such as IL-1B, IL-2, and IL-8 and displayed significant tumor-killing capacity. These results show that development of in vitro expanded UCB γδ T cell therapies is feasible. It could prove a valuable treatment modality for patients after umbilical cord blood transplantation