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Deployment and access to state-of-the-art diagnostic technologies remains a fundamental challenge in providing equitable global cancer care to low-resource settings. The expansion of digital pathology in recent years and its interface with computational biomarkers provides an opportunity to democratize access to personalized medicine. Here we describe a low-cost platform for digital side capture and computational analysis composed of open-source components. The platform provides low-cost ($200) digital image capture from glass slides and is capable of real-time computational image analysis using an open-source deep learning (DL) algorithm and Raspberry Pi ($35) computer. We validate the performance of deep learning models’ performance using images captured from the open-source workstation and show similar model performance when compared against significantly more expensive standard institutional hardware.

Ovarian cancer is characterized by early, diffuse metastatic spread with most women presenting with extensive abdominal metastasis at the time of diagnosis. Prior work demonstrated carcinoma-associated mesenchymal stem cells (CA-MSCs) enhance ovarian cancer metastasis through a process of direct cellular interaction and formation of heterocellular CA-MSC and tumor cell complexes. In this study, we demonstrated that CA-MSCs enhance metastasis by increasing tumor cell heterogeneity through mitochondrial donation. We showed that CA-MSCs directly interacted with ovarian cancer cells via tunneling nanotubules (TNTs), and CA-MSCs used these TNTs to transfer live mitochondria to adjacent ovarian cancer cells. This mitochondrial donation preferentially occurred with ovarian cancer cells that had the lowest mitochondrial mass, as quantified using live, actively respiring mitochondrial labeling. These ‘mito poor’ cancer cells demonstrated decreased proliferation, increased sensitivity to chemotherapy, and decreased oxidative phosphorylation compared to ‘mito rich’ cancer cells. CA-MSCs rescued the phenotypes of mito poor cancer cells, restoring their proliferative capacity, increasing chemotherapy resistance, and increasing oxidative phosphorylation. We validated these findings in a fully autologous system using CA-MSCs and cancer cells derived from the same patient to prevent confounding effects of cellular response to foreign organelle/DNA. Using a knockdown of the mitochondrial motor protein, MIRO1, in CA-MSCs, we demonstrated that mitochondrial transfer is necessary for the CA-MSC-mediated rescue of ‘mito poor’ cancer cells. Mitochondria of CA-MSC origin persisted in tumor cells over multiple passages. Importantly, CA-MSC mitochondrial donation occurred in vivo, significantly enhanced tumor cell heterogeneity and decreased survival in an orthotopic ovarian cancer mouse model. Collectively, this work identified CA-MSC mitochondrial transfer as a critical mediator of ovarian cancer cell survival, heterogeneity, and metastasis, and blocking CA-MSC mitochondrial transfer represents a unique therapeutic target in ovarian cancer.

Interleukin [IL]-23 is a member of the IL-12 family of cytokines and has been implicated in multiple inflammatory disorders including psoriasis, psoriatic arthritis, and the inflammatory bowel diseases [IBDs]. Blockade of both IL-12 and IL-23 using an antibody that targets a shared subunit is highly effective in treating psoriasis, and recent data suggest similar efficacy in IBD with minimal adverse events. In this review, we summarise published data on the efficacy of anti-IL-12/23 therapies in IBD as well as emerging data on more selective anti-IL-23 specific therapies. Last, we discuss novel therapeutics under development which target the IL-23 pathway in unique ways and suggest that a biomarker-driven approach will soon guide clinicians to prescribe anti-IL-23 therapies to the patients most likely to respond to them.

Human heart samples from the Sydney Heart Bank have become a de facto standard against which others can be measured. Crucially, the heart bank contains a lot of donor heart material: for most researchers this is the hardest to obtain and yet is necessary since we can only study the pathological human heart in comparison with a control, preferably a normal heart sample. It is not generally realised how important the control is for human heart studies. We review our studies on donor heart samples. We report the results obtained with 17 different donor samples collected from 1994 to 2011 and measured from 2005 to 2015 by our standard methodology for in vitro motility and troponin I phosphorylation measurements. The donor heart sample parameters are consistent between the hearts, over time and with different operators indicating that Sydney Heart Bank donor hearts are a valid baseline control for comparison with pathological heart samples. We also discuss to what extent donor heart samples are representative of the normal heart. Electronic supplementary material The online version of this article (10.1007/s12551-020-00740-2) contains supplementary material, which is available to authorized users.

This chapter describes the innovative use of a pre-existing framework from the field of Human–Computer Interaction to explore and reimagine reading spaces for beginner readers in primary schools in England. The chapter details the four phases of the research study, from a conceptual level to a physical outcome (a reading nook), the latter providing secondary findings about the value of secluded reading spaces for students in two English classrooms. In adapting and developing a design methodology within an educational context, it is hoped that this research will stimulate communication and dialogue between architects, educators, policy-makers and students. It also offers a contribution to the challenge of improving school design for pupils and practitioners by offering a framework through which education, specifically reading, can be viewed through the prism of design.

Staphylococcal peptidoglycan is characterized by pentaglycine cross-bridges that are cross-linked between adjacent wall peptides by penicillin-binding proteins to confer robustness and flexibility. In Staphylococcus aureus, pentaglycine cross-bridges are synthesized by three proteins: FemX adds the first glycine, and the homodimers FemA and FemB sequentially add two Gly-Gly dipeptides. Occasionally, serine residues are also incorporated into the cross-bridges by enzymes that have heretofore not been identified. Here, we show that the FemA/FemB homologues FmhA and FmhC pair with FemA and FemB to incorporate Gly-Ser dipeptides into cross-bridges and to confer resistance to lysostaphin, a secreted bacteriocin that cleaves the pentaglycine cross-bridge. FmhA incorporates serine residues at positions 3 and 5 of the cross-bridge. In contrast, FmhC incorporates a single serine at position 5. Serine incorporation also lowers resistance toward oxacillin, an antibiotic that targets penicillin-binding proteins, in both methicillin-sensitive and methicillin-resistant strains of S. aureus. FmhC is encoded by a gene immediately adjacent to lytN, which specifies a hydrolase that cleaves the bond between the fifth glycine of cross-bridges and the alanine of the adjacent stem peptide. In this manner, LytN facilitates the separation of daughter cells. Cell wall damage induced upon lytN overexpression can be alleviated by overexpression of fmhC. Together, these observations suggest that FmhA and FmhC generate peptidoglycan cross-bridges with unique serine patterns that provide protection from endogenous murein hydrolases governing cell division and from bacteriocins produced by microbial competitors.

Emma Dyer, previously a nursery manager, works with the charity MOVE, to support children who have physical disabilities to develop their movement opportunities via education. Here Emma discusses the importance of making early years settings more accessible for children with disabilities and ways in which this can be supported.

A 3-month-old baby is brought to the paediatric emergency department by their parents because of a fever. You decide to check their inflammatory markers. Their C-reactive protein (CRP) level comes back as 20 mg/L. Does this affect whether or not you start antibiotic therapy? Does it influence your decision to admit or discharge the patient? CRP is a commonly used biochemical test and yet its use is constantly debated and challenged. We look at the current evidence and suggest the best way to use this test in clinical practice.

Monogenic forms of diabetes (historically known as Maturity Onset Diabetes of the Young (MODY)) are caused by single gene mutations inherited in an autosomal dominant fashion that result in reduced pancreatic beta cell function. Children with these forms of diabetes may be misdiagnosed as having type 1 or 2 diabetes, which has important implications for treatment, genetic counselling, screening of family members and prognosis. Useful tools now exist to aid in their diagnosis and management. Here, we attempt to outline the clinical features that will help the physician make the differentiation from other diabetes subtypes.

A 13-year-old girl presents to the emergency department for the second time with an unresponsive episode. She has a GCS (Glasgow Coma Scale) score of 11 on arrival and all other observations are normal. The story is unclear, but there are ongoing safeguarding concerns and the family are known to social services. All investigations are normal. After a period of observation on the ward, her GCS returns to normal and she appears well. Both on the first presentation and this presentation ingestion of a toxin was suspected. However, this was denied by the patient and urine toxicology screen was negative. Does this rule out toxin ingestion? Will this change your management?

Mast cell tryptase (tryptase) is an enzyme produced almost exclusively by mast cells that is easy to measure using a widely available test. In this article we discuss the physiology of the mast cell and how that relates to IgE-mediated anaphylaxis and mastocytosis. We also describe the technical aspects of testing tryptase and the reported normal ranges in health. Finally we explore the diagnostic performance of serum mast cell tryptase measurements, when used to confirm anaphylaxis, estimate future anaphylaxis risk and in diagnosing/monitoring leukaemia.

There are over 300 new cases of imported paediatric malaria in the UK each year and this has been increasing over the last 20 years. Malaria in children is particularly difficult to diagnose because the initial presenting features are subtler than in adults and do not display the classical presenting features. However, they are also more likely to deteriorate rapidly and to develop severe malaria. The 'gold standard' for ruling out the diagnosis of malaria if clinically suspected is three negative thin and thick blood films, which require serial phlebotomy and the availability of trained technicians. There are now a range of other tests, including rapid diagnostic tests and PCR, as well as clinical features that make the diagnosis more or less likely. We explore the different tests available and whether these might replace the three negative blood films currently needed. We also look at whether we are able to use clinical features to aid the tests used for a diagnosis of imported malaria.

The relationship between school buildings and their pedagogies is complex. There is a consensus that the built environment through its structure and organisation can impact education, but it is often unclear or unknown how this happens in individual cases. Here we examine one design process, which led to the construction of a secondary school in England. Driven by an ethos of collaboration, it resulted in a structure that aimed to favour the students, and not the institution’s need for controlling them, and it supports collaborative project-based learning. We discuss in detail the design epistemology (’creative discipline’) of the architectural practice that facilitated this process, and the employed method (‘spin painting’ diagrams). We argue that this unique procedure was a catalyst for stimulating collective decision-making and the visual method employed was a tool for translating the pedagogical model into a tangible structure. It was invented specifically to facilitate generating a shared vision as a necessary condition of compatibility between the school’s educational approach and the building’s affordances for learning.

Spontaneous bleeding in the neonatal period is an unusual presentation, and yet one that can pose a challenge of both diagnosis and management to the general paediatrician. This case chronicles the diagnostic journey of an 8-day-old baby who presented with unrelenting bleeding from the umbilical cord and explores the clinical approach to bleeding in a neonate.

We generated a transgenic mouse model expressing the apical hypertrophic cardiomyopathy-causing mutation ACTC E99K at 50% of total heart actin and compared it with actin from patients carrying the same mutation. The actin mutation caused a higher Ca(2+) sensitivity in reconstituted thin filaments measured by in vitro motility assay (2.3-fold for mice and 1.3-fold for humans) and in skinned papillary muscle. The mutation also abolished the change in Ca(2+) sensitivity normally linked to troponin I phosphorylation. MyBP-C and troponin I phosphorylation levels were the same as controls in transgenic mice and human carrier heart samples. ACTC E99K mice exhibited a high death rate between 28 and 45 days (48% females and 22% males). At 21 weeks, the hearts of the male survivors had enlarged atria, increased interstitial fibrosis, and sarcomere disarray. MRI showed hypertrophy, predominantly at the apex of the heart. End-diastolic volume and end-diastolic pressure were increased, and relaxation rates were reduced compared with nontransgenic littermates. End-systolic pressures and volumes were unaltered. ECG abnormalities were present, and the contractile response to β-adrenergic stimulation was much reduced. Older mice (29-week-old females and 38-week-old males) developed dilated cardiomyopathy with increased end-systolic volume and continuing increased end-diastolic pressure and slower contraction and relaxation rates. ECG showed atrial flutter and frequent atrial ectopic beats at rest in some ACTC E99K mice. We propose that the ACTC E99K mutation causes higher myofibrillar Ca(2+) sensitivity that is responsible for the sudden cardiac death, apical hypertrophy, and subsequent development of heart failure in humans and mice.

Aims The vascular type of Ehlers–Danlos syndrome (EDS IV) is an autosomal-dominant disorder characterized by thin translucent skin and extensive bruising. Patients with EDS IV have reduced life expectancy (median 45–50 years) due to spontaneous rupture of arteries (particularly large arteries) or bowel. EDS IV results from mutation of the COL3A1 gene, which encodes the pro-α1 chains of type III collagen that is secreted into the extracellular matrix, e.g. by smooth muscle cells. A mouse model of EDS IV produced by targeted ablation of Col3a1 has been of limited use as only 10% of homozygous animals survive to adulthood, whereas heterozygous animals do not die from arterial rupture. We report a novel, exploitable model of EDS IV in a spontaneously generated mouse line. Methods and results Mice were identified by predisposition to sudden, unexpected death from dissection of the thoracic aorta. Aortic dissection inheritance was autosomal-dominant, presented at an early age (median, 6 weeks) with incomplete penetrance, and had a similar sex ratio bias as EDS IV (2:1, male:female). Molecular genetic analysis demonstrated that the causal mutation is a spontaneous 185 kb deletion, including the promoter region and exons 1–39, of the Col3a1 gene. As in EDS IV, aortic dissection was not associated with elevated blood pressure, aneurysm formation, or infection, but may result from aberrant collagen fibrillogenesis within the aortic wall. Conclusion This novel, exploitable mouse line that faithfully models the vascular aspects of human EDS IV provides an important new tool for advancing understanding of EDS IV and of aortic dissection in general.

In the past three years, Santos has undergone substantial growth across multiple jurisdictions. In this time the number of environmental specialists has more than doubled. Santos’ vision is to be a leading energy company for Australia and Asia. To do this we have to have the best people doing the right jobs. To support this, there are a number of approaches and tools that we have developed, including: Development planning systemsMailing lists and group emailsThe on-line Environment CommunityThe Environment Network meetingEnvironmental leaders meetingThe Environmental Excellence ForumDatabases A number of hurdles have been experienced along the path to developing Santos’ environmental professionals into a cohesive knowledgeable cohort. Challenges include: Changing regulatory and community expectationsShortage of appropriately skilled and experienced peopleChanging technologyBusy-nessChanging business needs The fundamental key to succeeding in developing a cohesive environmental community has been using consistent and regular communications. Santos’ journey to build an environmental profession made up of skilled and empowered people that know and support their colleagues is well underway. The challenge is to ensure that we can keep up as technology continues to evolve, community and regulators’ expectations change and the company strives to be a leading energy company for Australia and Asia.

We have investigated a transgenic mouse model of inherited dilated cardiomyopathy that stably expresses the ACTC E361G mutation at around 50% of total actin in the heart. F-actin isolated from ACTC E361G mouse hearts was incorporated into thin filaments with native human tropomyosin and troponin and compared with NTG mouse actin by in vitro motility assay. There was no significant difference in sliding speed, fraction of filaments motile or Ca(2+)-sensitivity (ratio EC(50) E361G/NTG=0.95+/-0.08). The Ca(2+)-sensitivity of force in skinned trabeculae from ACTC E361G mice was slightly higher than NTG (EC(50) E361G/NTG=0.78+/-0.04). The molecular phenotype was revealed when troponin was dephosphorylated; Ca(2+)-sensitivity of E361G-containing thin filaments was now lower than NTG (EC(50) E361G(dPTn)/NTG(dPTn)=2.15+/-0.09). We demonstrated that this was due to uncoupling of Ca(2+)-sensitivity from troponin I phosphorylation by comparing Ca(2+)-sensitivity of phosphorylated and dephosphorylated thin filaments. For NTG actin-containing thin filaments EC(50) native/dPTn=3.0+/-0.3 but for E361G-containing thin filaments EC(50) native/dPTn=1.04+/-0.07.We studied contractility in isolated myocytes and found no significant differences under basal conditions. We measured cardiac performance by cine-MRI, echocardiography and with a conductance catheter over a period of 4 to 18 months and found minimal systematic differences between NTG and ACTC E361G mice under basal conditions. However, the increase in septal thickening, ejection fraction, heart rate and cardiac output following dobutamine treatment was significantly less in ACTC E361G mice compared with NTG. We propose that the ACTC E361G mutation uncouples myofilament Ca(2+)-sensitivity from Troponin I phosphorylation and blunts the response to adrenergic stimulation, leading to a reduced cardiac reserve with consequent contractile dysfunction under stress, leading to dilated cardiomyopathy.

Telomerase is expressed in approximately 90% of breast cancers but not in normal somatic cells. The catalytic subunit of telomerase, human telomerase reverse transcriptase (hTERT), is a widely expressed tumour-associated antigen against which specific cytotoxic T lymphocyte (CTL) responses have been identified. Human CD8+ CTL have been shown to recognise hTERT synthetic peptides in an HLA-restricted manner and to lyse hTERT+ tumour cells. This study has investigated the specific CD8+ CTL response in a defined cohort of primary breast cancer patients against 3 hTERT peptides using an IFNgamma ELISpot assay. A specific CD8+ CTL response against at least one of the hTERT peptides was observed in 75% of cancer patients, with 8/17 patients responding to both HLA-A2 restricted peptides. In contrast, 38% of normal healthy female controls responded against a single peptide only. This suggests that a specific immune response exists against hTERT in breast cancer patients and that hTERT is a suitable target for tumour-based immunotherapy.

The mutation Gly99lys (E99K) in the cardiac actin (ACTC) gene has been found to cause hypertrophic cardiomyopathy (HCM) in 75 hypertrophic obstructive cardiomyopathy (HOCM) or LVNC patients. Transgenic mice expressing 50% E99K mutant cardiac actin in their hearts were generated and studied. Over 30% male and 70% female E99K mice died …

The mutation Gly99lys (E99K) in the cardiac actin (ACTC) gene was reported to cause Hypertrophic Cardiomyopathy in extensive clinical studies. Transgenic (TG) mice expressing 50% E99K mutant cardiac actin in their hearts were generated and studied. The mice show high mortality between 28 and 45 days old (70% females, 34% males).Thin filaments reconstituted with purified mouse f-actin from the survivors and human heart tropomyosin and troponin were studied by in vitro motility assay. The E99K thin filaments were 2.5 ± 0.6 times more Ca2+ sensitive than NTG thin filaments (p = 0.05). E99K actin thin filaments also exhibited a reduced response to troponin dephosphorylation (EC50 E99K/E99KdP = 1.1 ± 0.1 compared with 3.0 ± 0.3 for NTG /NTGdP).7 month-old E99K TG mice (n=9) and their NTG littermates (n=7) were studied using in vivo cine MRI. Abnormal cardiac morphology and significantly lower ejection fractions (56.5 vs. 65.2%) and reduced stroke volumes (26.0 vs. 42.3 μl) were observed in TG mice. Peak LV ejection rates were also reduced (188 ± 41 vs. 252 ± 49 μl/min). LV mass was similar between groups, but septal wall thickness was increased (1.5 vs. 1.0 mm).Left ventricular function of 9 month-old female E99K NTG (n=4) and TG (n=5) mice were studied with an in vivo conductance catheter. In TG mice ejection fraction was 20.2% less, end-diastolic pressure was 39.6% higher and relaxation rate was 50.0% slower.We conclude that the basic effect of E99K mutation is increased Ca2+-sensitivity and blunted response to troponin dephosphorylation and this leads to the high rate of sudden death at early ages, alterations to cardiac function and hypertrophy as observed in patients with hypertrophic cardiomyopathy.Supported by a grant from the British Heart Foundation