SETD1A-dependent EME1 transcription drives PARPi sensitivity in HR deficient tumour cells.

Background: Cells deficient in DNA repair factors breast cancer susceptibility 1/2 (BRCA1/2) or ataxia-telangiectasia mutated (ATM) are sensitive to poly-ADP ribose polymerase (PARP) inhibitors. Building on our previous findings, we asked how the lysine methyltransferase SETD1A contributed to PARP inhibitor-mediated cell death in these contexts and determined the mechanisms responsible.

Methods: We used cervical, breast, lung and ovarian cancer cells bearing mutations in BRCA1 or ATM and depleted SETD1A using siRNA or CRISPR/Cas9.

CLEAR - clozapine in early psychosis: study protocol for a multi-centre, randomised controlled trial of clozapine vs other antipsychotics for young people with treatment resistant schizophrenia in real world settings.

Background: Clozapine is an antipsychotic drug with unique efficacy, and it is the only recommended treatment for treatment-resistant schizophrenia (TRS: failure to respond to at least two different antipsychotics). However, clozapine is also associated with a range of adverse effects which restrict its use, including blood dyscrasias, for which haematological monitoring is required. As treatment resistance is recognised earlier in the illness, the question of whether clozapine should be prescribed in children and young people is increasingly important.

MYBL2 amplification in breast cancer: Molecular mechanisms and therapeutic potential.

The MYBL2 gene, also known as B-MYB, is essential to regulate vital cellular processes including cell proliferation, differentiation and DNA repair. Changes in these pathways can facilitate cancer development and as such targeting these processes represent an effective method to treat multiple cancer types. Alterations in gene expression have been identified in cancer cells including changes in MYBL2, which appears to be of particular significance in breast cancer (BC) patients.

Dependence on Myb expression is attenuated in myeloid leukaemia with N-terminal CEBPA mutations.

Mutations at the N- or C-terminus of C/EBPα are frequent in acute myeloid leukaemia (AML) with normal karyotype. Here, we investigate the role of the transcription factor Myb in AMLs driven by different combinations of CEBPA mutations. Using knockdown of Myb in murine cell lines modelling the spectrum of CEBPA mutations, we show that the effect of reduced Myb depends on the mutational status of the two Cebpa alleles.

MYBL2 Supports DNA Double Strand Break Repair in Hematopoietic Stem Cells.

Myelodysplastic syndromes (MDS) are a heterogeneous group of diseases characterized by blood cytopenias that occur as a result of somatic mutations in hematopoietic stem cells (HSC). MDS leads to ineffective hematopoiesis, and as many as 30% of patients progress to acute myeloid leukemia (AML). The mechanisms by which mutations accumulate in HSC during aging remain poorly understood.

The productivity limit of manufacturing blood cell therapy in scalable stirred bioreactors.

Manufacture of red blood cells (RBCs) from progenitors has been proposed as a method to reduce reliance on donors. Such a process would need to be extremely efficient for economic viability given a relatively low value product and high (2 × 10 ) cell dose. Therefore, the aim of these studies was to define the productivity of an industry standard stirred-tank bioreactor and determine engineering limitations of commercial red blood cells production.

The autoimmune-associated genetic variant PTPN22 R620W enhances neutrophil activation and function in patients with rheumatoid arthritis and healthy individuals.

Objectives: A genetic variant of the leukocyte phosphatase PTPN22 (R620W) is strongly associated with autoimmune diseases including rheumatoid arthritis (RA). Functional studies on the variant have focussed on lymphocytes, but it is most highly expressed in neutrophils. We have investigated the effects of the variant on neutrophil function in health and in patients with RA.

Differential expression of CD148 on leukocyte subsets in inflammatory arthritis.

Introduction: Monocytic cells play a central role in the aetiology of rheumatoid arthritis, and manipulation of the activation of these cells is an approach currently under investigation to discover new therapies for this and associated diseases. CD148 is a transmembrane tyrosine phosphatase that is highly expressed in monocytes and macrophages and, since this family of molecules plays an important role in the regulation of cell activity, CD148 is a potential target for the manipulation of macrophage activation. For any molecule to be considered a therapeutic target, it is important for it to be increased in activity or expression during disease.

Does oxidative inactivation of CD45 phosphatase in rheumatoid arthritis underlie immune hyporesponsiveness?

The protein tyrosine phosphatase (PTP) CD45 is critical in regulating the earliest steps in T-cell-receptor signaling but, similar to all PTPs, it is susceptible to oxidative inactivation. Given the widely reported effects of oxidant damage associated with rheumatoid arthritis (RA), we examined whether CD45 phosphatase activity was altered in CD4(+) T cells from RA patients and related this to CD4(+) T-cell function and redox status. CD45 phosphatase specific activity in T cells from RA peripheral blood (PB) and synovial fluid was 56% and 59% lower than in healthy control (HC) PB, respectively.

Measuring the specific activity of the protein tyrosine phosphatase Lyp.

Altered function of the protein tyrosine phosphatase (PTP) Lyp (PTPN22) has been implicated in the pathogenesis of a number of human diseases, and so accurate assessment of its functional activity is needed to further our understanding of its biology. We have developed an in vitro method to measure the specific catalytic activity of the Lyp phosphatase. Lyp is captured from cell lysates using an anti-Lyp monoclonal antibody coated 96-well plate, and activity measured by dephosphorylation of a fluorescent substrate, 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP).

Impact of ambulance crew configuration on simulated cardiac arrest resuscitation.

Background: Despite the widespread use of both two paramedic and single paramedic ambulance crews, there is little evidence regarding differences between these two staffing configurations in the delivery of patient care.

Objectives: To determine potential differences in care provided by each of these ambulance configurations in the resuscitation of a cardiac arrest victim in ventricular fibrillation.

Methods: Fifteen paramedic-paramedic and 15 paramedic-EMT crews were recruited to perform resuscitation on a high-fidelity human simulator (Laerdal SimMan).

Radiation dosimetry by automatic image analysis of dicentric chromosomes.

A system for scoring dicentric chromosomes by image analysis comprised fully automatic location of mitotic cells, automatic retrieval, focus and digitization at high resolution, automatic rejection of nuclei and debris and detection and segmentation of chromosome clusters, automatic centromere location, and subsequent rapid interactive visual review of potential dicentric chromosomes to confirm positives and reject false positives. A calibration set of about 15,000 cells was used to establish the quadratic dose response for 60Co gamma-irradiation. The dose-response function parameters were established by a maximum likelihood technique, and confidence limits in the dose response and in the corresponding inverse curve, of estimated dose for observed dicentric frequency, were established by Monte Carlo techniques.

Automated densitometry of cell populations in a continuous-motion imaging cell scanner.

Continuous-motion imaging provides a method for the rapid quantitative analysis of slide-mounted cell preparations. Densitometric and morphometric cellular features can be measured and used to classify individual cells. Tests on the densitometric accuracy of the CERVIFIP CMI system show a c.

A fast interval processor (FIP) for cervical prescreening.

A cervical prescreening device (FIP: fast interval processor) designed to scan and classify a slide-mounted specimen within two minutes is described. The image analysis techniques are based directly on the MRC Cerviscan equipment with the minimal conversion needed to adapt these techniques for interval processing. A high scanning rate is achieved by scanning with a charge-coupled diode linear image sensor along one axis and by stepping the microscope stage continuously along the other axis.

A cytogeneticist's microscope.

It is demonstrated that there are a number of advantages in using a mechanised microscope for scoring a large number of metaphase cells from human blood lymphocyte preparations. Following the development of an automatic metaphase spread finding machine based upon a large motorised microscope and a synchronous closed circuit television camera and flashing light source, a much smaller machine which is more appropriate to the cytogenetics laboratory, but with a similar metaphase finding performance has been constructed. The new machine which consists of a Cambridge Instruments 1 micron stepping microscope stage, a linear diode array scanner and a computer is described in detail.