Regulation of CD38 by IRF4 in chronic lymphocytic leukemia

PhD ThesisGenome‐wide association analysis identified rs872071, a common variant in the 3’UTR of IRF4, as tagging a risk allele for chronic lymphocytic leukaemia (CLL). The risk allele is significantly associated with expression of CD38, a poor prognostic marker in CLL. IRF4 is a transcription factor with pleiotropic roles in the regulation of B cell development, and interrogation of the CD38 gene identified a number of putative binding sites for IRF4, suggesting that CD38 may be transcriptionally regulated by IRF4. Chromatin immunoprecipitation (ChIP) demonstrated significant IRF4‐CD38 binding at two composite ETS/IRF consensus element (EICE) sites in SU‐DHL‐6 and MEC‐1 mature B cell lines. Furthermore, there was evidence of IRF4‐CD38 binding at these EICE sites in primary CLL lymphocytes in some, but not all, cases. ChIP studies using markers of histone methylation suggested increased transcriptional activation at the IRF4‐CD38 binding sites in MEC‐1 cells with IRF4 knockdown. In contrast, transcriptional activity at these sites appeared to be reduced in SU‐DHL‐6 cells with IRF4 knockdown. Co‐culture of primary CLL lymphocytes on a CD40L‐expressing monolayer led to an upregulation of IRF4 expression, but no consistent effect on CD38 expression. In addition, ChIP studies using markers of histone methylation were suggestive of reduced transcriptional activity at the IRF4‐CD38 binding site after CD40L co‐culture, suggesting that IRF4 may be a negative regulator of CD38 in CLL. Taken together, the evidence indicates that IRF4 binds to the CD38 locus. However, direct experimental evidence for an effect on CD38 expression is lacking. It is necessary to consider that the interaction between IRF4 and CD38 is unlikely to be a linear signal transduction pathway. Indeed the prevailing evidence regarding the function of IRF4 in B cells suggests a complex signalling network involving numerous other transcription factors and cytokines. Furthermore, IRF4 has been suggested as a putative therapeutic target in haematological malignancies including myeloma. Transient IRF4 knockdown using RNA ii interference (RNAi) techniques was tolerated by SU‐DHL‐6, MEC‐1 and lymphoblastoid TK6 cell lines, though cell proliferation in TK6 and SU‐DHL‐6 was significantly impaired. Long‐term stable knockdown was also tolerated in TK6 cells, but not in MEC‐1 cells, suggesting an essential role for IRF4 in maintenance of MEC‐1 cells. Targeted knockdown of IRF4 also sensitised TK6 B cells and MEC‐1 CLL cells to the growth inhibitory effects of fludarabine, a nucleoside analogue used in the treatment of CLL. These findings indicate that IRF4 is necessary to the maintenance of MEC‐1 cells, which represent a CLL cell line model. A greater understanding of the role of IRF4 in the development and maintenance of CLL may indicate new therapeutic targets in CLL. Indeed, a recently developed novel mouse model of CLL has indicated that deficient IRF4 expression predisposes to the development of CLL. However, the role of IRF4 in the maintenance of CLL cells is yet to be determined. The heterogeneous nature of IRF4 expression in primary CLL lymphocytes determined here, and its potentially pleiotropic role in the transcriptional regulation of CD38 in B cells at different stages of differentiation, suggest that if IRF4 is a key player in the maintenance of the leukaemic clone, its role may be context or patient specific, and dependent on other signalling components or somatic genetic abnormalities.Bright Red and the Medical Research Counci

Creative approaches to dark skies research: a dialogue between two artist-researchers

Though the arts are vital in engaging our imaginations of the night sky, the role and value of arts-based approaches to dark sky research remains underexplored. Our collaborative chapter addresses this through a dialogue between two artist-researchers, whose PhD research is produced in partnership with two UK-based international dark sky parks (IDSPs). Reflecting on our experiences, insights, and challenges, this chapter offers a unique perspective on the creative process in dark sky contexts. We explore how arts-based research can enhance the recreational activity taking place under dark skies and inform practices on the ground whilst also inviting IDSP stakeholders to engage in the creation of novel dark sky research. The chapter closes with a set of critical reflections and propositions for setting up future opportunities for arts-based knowledge creation in dark sky contexts

Assessing Quality Simulated Participant Programs: A Case Study of Bond University’s Simulated Participant Program

Assessing the quality of Simulated Participant Programs (SPPs) has long been a challenge for health professions faculty. A number of frameworks exist to determine markers of quality in SPPs, such as the Association of Standardised Patient Educators (ASPE) Standards of Best Practice (SOBP) and the UK Simulated Patient Common Framework (Lewis et al, 2017; Gough et al., 2015). Both frameworks present domains for assessing quality in the areas of working environments, program management, training, risk assessment and other quality assurance issues. This abstract presents the review of the Bond UniversitySimulated Participant Program in 2021 and 2022, which utilised the UK Simulated Patient Common Framework.ResultsOver the course of the review, the Bond University Participant Program demonstrated significant improvement between the first and second reviews. In 2021, the 5 domains (Resource considerations, recruitment and selection process, training requirements, risk assessments and quality assurance procedures) were either not met or partially met. In 2022, 4 out of 5 domains were met, with one area remaining partially met - challenges remain with regards to issues of ensuring staff diversity and minimising stereotypes.ConclusionsThe framework was a useful tool for evaluating the quality benchmarks of an SPP, however, there were gaps in the framework that present opportunities for the simulation and health professions community to explore

Dynamic clonal progression in xenografts of acute lymphoblastic leukemia with intrachromosomal amplification of chromosome 21

Intrachromosomal amplification of chromosome 21 is a heterogeneous chromosomal rearrangement occurring in 2% of childhood precursor B-cell acute lymphoblastic leukemia. There are no cell lines with iAMP21 and these abnormalities are too complex to faithfully engineer in animal models. As a resource for future functional and pre-clinical studies, we have created xenografts from intrachromosomal amplification of chromosome 21 leukemia patient blasts and characterised them by in-vivo and ex-vivo luminescent imaging, FLOW immunophenotyping, and histological and ultrastructural analysis of bone marrow and the central nervous system. Investigation of up to three generations of xenografts revealed phenotypic evolution, branching genomic architecture and, compared with other B-cell acute lymphoblastic leukemia genetic subtypes, greater clonal diversity of leukemia initiating cells. In support of intrachromosomal amplification of chromosome 21 as a primary genetic abnormality, it was always retained through generations of xenografts, although we also observed the first example of structural evolution of this rearrangement. Clonal segregation in xenografts revealed convergent evolution of different secondary genomic abnormalities implicating several known tumour suppressor genes and a region, containing the B-cell adaptor, PIK3AP1, and nuclear receptor co-repressor, LCOR, in the progression of B-ALL. Tracking of mutations in patients and derived xenografts provided evidence for co-operation between abnormalities activating the RAS pathway in B-ALL and for their aggressive clonal expansion in the xeno-environment. Bi-allelic loss of the CDKN2A/B locus was recurrently maintained or emergent in xenografts and also strongly selected as RNA sequencing demonstrated a complete absence of reads for genes associated with the deletions

Ecological and temperature controls on Mg/Ca ratios of Globigerina bulloides from the southwest Pacific Ocean

We present Mg/Ca data for Globigerina bulloides from 10 core top sites in the southwest Pacific Ocean analyzed by laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). Mg/Ca values in G. bulloides correlate with observed ocean temperatures (7 degrees C-19 degrees C), and when combined with previously published data, an integrated Mg/Ca-temperature calibration for 7 degrees C-31 degrees C is derived where Mg/Ca (mmol/mol) = 0.955 x e(0.068 x T) (r(2) = 0.95). Significant variability of Mg/Ca values (20%-30%) was found for the four visible chambers of G. bulloides, with the final chamber consistently recording the lowest Mg/Ca and is interpreted, in part, to reflect changes in the depth habitat with ontogeny. Incipient and variable dissolution of the thin and fragile final chamber, and outermost layer concomitantly added to all chambers, caused by different cleaning techniques prior to solution-based ICPMS analyses, may explain the minor differences in previously published Mg/Ca-temperature calibrations for this species. If the lower Mg/Ca of the final chamber reflects changes in depth habitat, then LA-ICPMS of the penultimate (or older) chambers will most sensitively record past changes in near-surface ocean temperatures. Mean size-normalized G. bulloides test weights correlate negatively with ocean temperature (T = 31.8 x e(-30.5xwtN); r(2) = 0.90), suggesting that in the southwest Pacific Ocean, temperature is a prominent control on shell weight in addition to carbonate ion levels

The Interfascicular Matrix of Energy Storing Tendons Houses Heterogenous Cell Populations Disproportionately Affected by Aging

Energy storing tendons such as the human Achilles and equine superficial digital flexor tendon (SDFT) are prone to injury, with incidence increasing with aging, peaking in the 5th decade of life in the human Achilles tendon. The interfascicular matrix (IFM), which binds tendon fascicles, plays a key role in energy storing tendon mechanics, and aging alterations to the IFM negatively impact tendon function. While the mechanical role of the IFM in tendon function is well-established, the biological role of IFM-resident cell populations remains to be elucidated. Therefore, the aim of this study was to identify IFM-resident cell populations and establish how these populations are affected by aging. Cells from young and old SDFTs were subjected to single cell RNA-sequencing, and immunolabelling for markers of each resulting population used to localise cell clusters. Eleven cell clusters were identified, including tenocytes, endothelial cells, mural cells, and immune cells. One tenocyte cluster localised to the fascicular matrix, whereas nine clusters localised to the IFM. Interfascicular tenocytes and mural cells were preferentially affected by aging, with differential expression of genes related to senescence, dysregulated proteostasis and inflammation. This is the first study to establish heterogeneity in IFM cell populations, and to identify age-related alterations specific to IFM-localised cells

Financial incentives for large-scale wetland restoration: beyond markets to common asset trusts

Wetlands provide $47.4 trillion/year worth of ecosystem services globally and support immense biodiversity, yet face widespread drainage and pollution, and large-scale wetlands restoration is urgently needed. Payment for ecosystem service (PES) schemes provide a viable avenue for funding large-scale wetland restoration. However, schemes around the globe differ substantially in their goals, structure, challenges, and effectiveness in supporting large-scale wetland restoration. Here, we suggest wetland-based PES schemes use common asset trusts (CATs) to build investment portfolios of wetlands across landscapes that sustain and enhance overall provision of multiple ecosystem services. CATs can meet the needs of multiple investors, permit bundled payments, and provide flexibility to invest in the restoration of numerous services/values, all using a coordinated, highly collaborative, prioritized, and transparent process. CATs would support financial viability, facilitate efficiency to reduce administrative burdens, and enable credibility and social licence building to restore wetland values and services globally

The Mpumalanga Men's Study (MPMS): results of a baseline biological and behavioral HIV surveillance survey in two MSM communities in South Africa

The Mpumalanga Men's Study (MPMS) is the assessment of the Project Boithato HIV prevention intervention for South African MSM. Boithato aims to increase consistent condom use, regular testing for HIV-negative MSM, and linkage to care for HIV-positive MSM. The MPMS baseline examined HIV prevalence and associated risk behaviors, and testing, care, and treatment behaviors among MSM in Gert Sibande and Ehlanzeni districts in Mpumalanga province, South Africa in order to effectively target intervention activities. We recruited 307 MSM in Gert Sibande and 298 in Ehlanzeni through respondent-driven sampling (RDS) between September 2012-March 2013. RDS-adjusted HIV prevalence estimates are 28.3% (95% CI 21.1%-35.3%) in Gert Sibande, and 13.7% (95% CI 9.1%-19.6%) in Ehlanzeni. Prevalence is significantly higher among MSM over age 25 [57.8% (95% CI 43.1%-72.9%) vs. 17.9% (95% CI 10.6%-23.9%), P <0.001 in Gert Sibande; 34.5% (95%CI 20.5%-56.0%) vs. 9.1% (95% CI 4.6%-13.9%), P <0.001 in Ehlanzeni]. In Gert Sibande, prevalence is higher among self-identified gay and transgender MSM vs. other MSM [39.3% (95%CI, 28.3%-47.9%), P <0.01], inconsistent condom users [38.1% (18.1%-64.2%), P <0.05], those with a current regular male partner [35.0% (27.1%-46.4%), P<0.05], and those with lifetime experience of intimate partner violence with men [40.4%, (95%CI 28.9%-50.9%), P <0.05]. Prevalence of previous HIV testing was 65.8% (95%CI 58.8%-74.0%) in Gert Sibande, and 69.3% (95%CI 61.9%-76.8%) in Ehlanzeni. Regular HIV testing was uncommon [(34.6%, (95%CI 27.9%-41.4%) in Gert Sibande; 31.0% (95%CI 24.9%-37.8%) in Ehlanzeni]. Among HIV-positive participants, few knew their status (28.1% in Gert Sibande and 14.5% in Ehlanzeni), or were appropriately linked to care (18.2% and 11.3%, respectively), or taking antiretroviral therapy (13.6% and 9.6% respectively). MPMS results demonstrate the importance of implementing interventions for MSM to increase consistent condom use, regular HIV testing, and linkage and engagement in care for HIV-infected MSM

Current Trends of Infectious Complications following Hematopoietic Stem Cell Transplantation in a Single Center

This study was to analyze the infectious complications after hematopoietic stem cell transplantation (HSCT) according to the recent changes of HSCT. Medical records of 379 adult patients who underwent HSCT consecutively at Catholic HSCT Center from January 2001 to December 2002 were reviewed retrospectively. Allogeneic HSCT accounted for 75.7% (287/379) and autologous HSCT for 24.3% (92/379). During pre-engraftment period, bacterial infection was predominant, and E. coli was still the most common organism. After engraftment, viral infection was predominant. The incidence of invasive fungal infection showed bimodal distribution with peak correlated with neutropenia and graft-versus-host disease (GVHD). The overall mortality and infection-related mortality rates according to 3 periods were as follows; during pre-engraftment, 3.16% (12/379) and 1.8% (7/379); during midrecovery period, 7.9% (29/367) and 4.1% (15/367); during late-recovery period, 26.9% (91/338), and 15.9% (54/338). Risk factors for infection-related mortality were as follows; during pre-engraftment period, fungal infection and septic shock; during the mid-recovery period, hemorrhagic cystitis and delayed engraftment; during the late-recovery period, fungal infection, chronic GVHD, and relapse. In conclusion, infection was still one of the main complications after HSCT and highly contributes to mortality. The early diagnosis and the effective vaccination strategy are needed for control of infections