The complex regulatory mechanisms of the BCL-2 family of proteins in cancer

Resistance to apoptosis is a key hallmark of most cancers. The BCL-2 family of proteins are the major arbiters of the cell death program and the interactions between pro- and anti-apoptotic members of this family are believed to shift the fate of a cell towards life or death. The anti-apoptotic BCL-2 family proteins (BCL-2, BCL-XL, MCL-1, BCL-w and BCL-2A1) are often overexpressed in cancer, thereby promoting abnormal cell survival. As a result, these proteins are highly attractive targets for novel chemotherapeutic approaches. The recent clinical success of inhibitors against BCL-2 (Navitoclax and Venetoclax) has launched the compounds known as BH3 mimetics into the chemotherapeutic spotlight and has led to the successful development of several highly potent MCL-1 inhibitors. The targeting of MCL-1 in cancer is critical to restoring sensitivity to other chemotherapeutic approaches, as MCL-1 is strongly associated with abnormal cell survival and chemoresistance in many cancers. The aims of this study were to (1) assess the specificity and potency of the novel MCL-1 inhibitor, S63845, as a single agent and in conjunction with other BH3 mimetics, to induce apoptosis in a range of cancer cell lines (2) characterise the dependence on BH3-only members for apoptosis induced by BH3 mimetics and (3) identify novel MCL-1 interacting partners which could explain BH3-only protein-independent apoptosis. Using a panel of cancer cell lines with various dependencies on BCL-2 family members, it was shown that S63845 is an extremely potent MCL-1 inhibitor which can induce extensive apoptosis alone or in combination with other BH3 mimetics. Immunoprecipitation studies of the BCL-2 family during BH3 mimetic-induced apoptosis revealed that the interactions of a pro-survival protein with certain BH3-only proteins might not exclusively dictate sensitivity to certain BH3 mimetics. Moreover, extensive study of cells lacking the 8 key BH3-only proteins showed that BH3 mimetics can induce a BAX-dependent but BH3-only protein-independent apoptosis, going against the accepted dogma of BH3 mimetic action. With the possibility of other BH3-only-like proteins existing to mediate the apoptosis seen in the absence of the 8 major BH3-only proteins, novel interacting partners of MCL-1 were explored by mass spectrometry. DRP-1, a mitochondrial fission regulator, was thus identified as an MCL-1-interacting protein. While knockdown of DRP-1 could abrogate BH3 mimetic-induced apoptosis in H1299 cells, it did not interact with MCL-1 via a BH3 motif and exacerbated apoptosis in the cells lacking the BH3-only proteins. Thus, DRP-1, while confirmed as a regulator of apoptosis, was not proposed to function like a novel BH3-only protein which can activate BAX/BAK in the absence of the BH3-only proteins. Overall this study reveals that S63845 is an important and highly effective MCL-1 inhibitor with strong potential for use in the clinic both as a monotherapy and in combination with other approaches. Moreover, the mechanism underlying BH3 mimetic-induced apoptosis and BAX activation has been revealed to be much more complicated than currently known, as it appears that the BH3-only proteins are dispensable for apoptosis following pro-survival protein neutralisation. Future studies should clarify how BAX can initiate MOMP without the BH3-only proteins, be it through activation of BAX by the outer mitochondrial membrane directly or, perhaps, the existence of novel BH3-only-like proteins which can activate BAX independently of the 8 key BH3-only proteins

Quantifying the effect of population mixing on childhood leukaemia risk: the Seascale cluster

A statistical model was developed based on Poisson regression of incidence of childhood leukaemia and non-Hodgkin’s lymphoma (NHL) in relation to population mixing among all 119 539 children born 1969–1989 to mothers living in Cumbria, north-west England, (excluding Seascale). This model was used to predict the number of cases in Seascale (the village adjacent to the Sellafield nuclear installation) children, born 1950–1989 and diagnosed before 1993. After allowing for age, the incidence of acute lymphoblastic leukaemia (ALL) and NHL was significantly higher among children born in areas with the highest levels of population mixing, relative risk (RR) = 11.7 (95% confidence interval (CI) 3.2–43) and was highest among children of incomers. The model predicted up to 3.0 (95% CI 1.3–6.0) cases of ALL/NHL in children born in Seascale compared to six observed and 2.0 (95% CI 1.0–3.4) cases in children resident, but not born, in Seascale compared to two observed. Population mixing is a significant risk factor for ALL/NHL, especially in young children, accounting for over 50% of cases in Cumbria and most cases in Seascale. © 1999 Cancer Research Campaig

Carbon storage and DNA absorption in allophanic soils and paleosols

Andisols and andic paleosols dominated by the nanocrystalline mineral allophane sequester large amounts of carbon (C), attributable mainly to its chemical bonding with charged hydroxyl groups on the surface of allophane together with its physical protection in nanopores within and between allophane nanoaggregates. C near-edge X-ray absorption fine structure (NEXAFS) spectra for a New Zealand Andisol (Tirau series) showed that the organic matter (OM) mainly comprises quinonic, aromatic, aliphatic, and carboxylic C. In different buried horizons from several other Andisols, C contents varied but the C species were similar, attributable to pedogenic processes operating during developmental upbuilding, downward leaching, or both. The presence of OM in natural allophanic soils weakened the adsorption of DNA on clay; an adsorption isotherm experiment involving humic acid (HA) showed that HA-free synthetic allophane adsorbed seven times more DNA than HA-rich synthetic allophane. Phosphorus X-ray absorption near-edge structure (XANES) spectra for salmonsperm DNA and DNA-clay complexes indicated that DNA was bound to the allophane clay through the phosphate group, but it is not clear if DNA was chemically bound to the surface of the allophane or to OM, or both. We plan more experiments to investigate interactions among DNA, allophane (natural and synthetic), and OM. Because DNA shows a high affinity to allophane, we are studying the potential to reconstruct late Quaternary palaeoenvironments by attempting to extract and characterise ancient DNA from allophanic paleosol

The JCMT Legacy Survey of the Gould Belt: Mapping 13CO and C 18O in Orion A

The Gould Belt Legacy Survey will map star-forming regions within 500 pc, using Heterodyne Array Receiver Programme (HARP), Submillimetre Common-User Bolometer Array 2 (SCUBA-2) and Polarimeter 2 (POL-2) on the James Clerk Maxwell Telescope (JCMT). This paper describes HARP observations of the J= 3 → 2 transitions of 13CO and C18O towards Orion A. The 15 arcsec resolution observations cover 5 pc of the Orion filament, including OMC 1 (including BN–KL and Orion bar), OMC 2/3 and OMC 4, and allow a comparative study of the molecular gas properties throughout the star-forming cloud. The filament shows a velocity gradient of ∼1 km s−1 pc−1 between OMC 1, 2 and 3, and high-velocity emission is detected in both isotopologues. The Orion Nebula and Bar have the largest masses and linewidths, and dominate the mass and energetics of the high-velocity material. Compact, spatially resolved emission from CH3CN, 13CH3OH, SO, HCOOCH3, CH3CHO and CH3OCHO is detected towards the Orion Hot Core. The cloud is warm, with a median excitation temperature of ∼24 K; the Orion Bar has the highest excitation temperature gas, at >80 K. The C18O excitation temperature correlates well with the dust temperature (to within 40 per cent). The C18O emission is optically thin, and the 13CO emission is marginally optically thick; despite its high mass, OMC 1 shows the lowest opacities. A virial analysis indicates that Orion A is too massive for thermal or turbulent support, but is consistent with a model of a filamentary cloud that is threaded by helical magnetic fields. The variation of physical conditions across the cloud is reflected in the physical characteristics of the dust cores. We find similar core properties between starless and protostellar cores, but variations in core properties with position in the filament. The OMC 1 cores have the highest velocity dispersions and masses, followed by OMC 2/3 and OMC 4. The differing fragmentation of these cores may explain why OMC 1 has formed clusters of high-mass stars, whereas OMC 4 produces fewer, predominantly low-mass stars

Exploratory Investigation of Head Stability in Children with Cerebral Palsy and Typically Developing Children during a Targeted Stepping Task

Children with cerebral palsy (CP) exhibit head instability during simple overground walking, which may comprise sensory input and reduce stepping accuracy. Investigations of head stability during more challenging tasks, where fall risk may be increased, are limited. This study explored differences in head stability between ambulatory children with hemiplegic CP (N = 9) and diplegia (N = 9) (GMFCS I and II) and typically developing (TD) children (N = 8) during a targeted stepping task. All children completed five trials stepping into two successive rectangular floor-based targets whilst walking along an 8 m walkway. Three-dimensional motion capture enabled calculation of head stability and foot placement within and before each target. A two-way mixed-design ANOVA compared differences between all groups and target approach. Children with diplegic CP showed greater sagittal, frontal, and resultant head-to-laboratory and head-to-trunk head instability compared to children with hemiplegic CP and TD children. Anteroposterior foot placement error was significantly greater in children with hemiplegic CP (8.5 ± 5.0 cm) compared to TD children (3.8 ± 1.5 cm). Group differences in head instability were not consistent with group differences in foot placement error. To better understand how head instability might affect fall risk in children with CP, more challenging environments should be tested in future

Higher risk for acute childhood lymphoblastic leukaemia in Swedish population centres 1973-94

A population-based sample of acute childhood leukaemia cases in Sweden 1973–94 was analysed by a geographical information system (GIS) for spatial leukaemia distribution in relation to population density. The annual incidence rate for acute lymphoblastic leukaemia (ALL) was 3.6, and for acute non-lymphoblastic leukaemia (ANLL) 0.7, cases per 100 000 children. Incidence rates in population centres, constituting 1.3% of Sweden's land area and approximately 80% of the population, compared with the rest of Sweden showed a statistically significant excess of ALL [odds ratio (OR) 1.68; 95% confidence interval (CI) 1.44–1.95], but not ANLL (OR 1.13; 95% CI 0.98–1.32). An increasing trend, however not statistically significant, was found for ALL incidence with both increasing population density in parishes and increasing degree of urbanity in municipalities. These findings support the theories that some environmental factors associated with high population density, such as infectious agents, may be of aetiological importance for childhood acute lymphoblastic leukaemia. © 1999 Cancer Research Campaig

Nitrogen and sulphur management: challenges for organic sources in temperate agricultural systems

A current global trend towards intensification or specialization of agricultural enterprises has been accompanied by increasing public awareness of associated environmental consequences. Air and water pollution from losses of nutrients, such as nitrogen (N) and sulphur (S), are a major concern. Governments have initiated extensive regulatory frameworks, including various land use policies, in an attempt to control or reduce the losses. This paper presents an overview of critical input and loss processes affecting N and S for temperate climates, and provides some background to the discussion in subsequent papers evaluating specific farming systems. Management effects on potential gaseous and leaching losses, the lack of synchrony between supply of nutrients and plant demand, and options for optimizing the efficiency of N and S use are reviewed. Integration of inorganic and organic fertilizer inputs and the equitable re-distribution of nutrients from manure are discussed. The paper concludes by highlighting a need for innovative research that is also targeted to practical approaches for reducing N and S losses, and improving the overall synchrony between supply and demand

Population genetics of cancer cell clones: possible implications of cancer stem cells

Abstract Background The population dynamics of the various clones of cancer cells existing within a tumour is complex and still poorly understood. Cancer cell clones can be conceptualized as sympatric asexual species, and as such, the application of theoretical population genetics as it pertains to asexual species may provide additional insights. Results The number of generations of tumour cells within a cancer has been estimated at a minimum of 40, but high cancer cell mortality rates suggest that the number of cell generations may actually be in the hundreds. Such a large number of generations would easily allow natural selection to drive clonal evolution assuming that selective advantages of individual clones are within the range reported for free-living animal species. Tumour cell clonal evolution could also be driven by variation in the intrinsic rates of increase of different clones or by genetic drift. In every scenario examined, the presence of cancer stem cells would require lower selection pressure or less variation in intrinsic rates of increase. Conclusions The presence of cancer stem cells may result in more rapid clonal evolution. Specific predictions from theoretical population genetics may lead to a greater understanding of this process.</p

Analytic philosophy for biomedical research: the imperative of applying yesterday's timeless messages to today's impasses

The mantra that "the best way to predict the future is to invent it" (attributed to the computer scientist Alan Kay) exemplifies some of the expectations from the technical and innovative sides of biomedical research at present. However, for technical advancements to make real impacts both on patient health and genuine scientific understanding, quite a number of lingering challenges facing the entire spectrum from protein biology all the way to randomized controlled trials should start to be overcome. The proposal in this chapter is that philosophy is essential in this process. By reviewing select examples from the history of science and philosophy, disciplines which were indistinguishable until the mid-nineteenth century, I argue that progress toward the many impasses in biomedicine can be achieved by emphasizing theoretical work (in the true sense of the word 'theory') as a vital foundation for experimental biology. Furthermore, a philosophical biology program that could provide a framework for theoretical investigations is outlined