Cellular stress triggers the human topoisomerase I damage response independently of DNA damage in a p53 controlled manner

The 'human topoisomerase I (htopoI) damage response' was reported to be triggered by various kinds of DNA lesions. Also, a high and persistent level of htopoI cleavage complexes correlated with apoptosis. In the present study, we demonstrate that DNA damage-independent induction of cell death using colcemid and tumor necrosis factor is also accompanied by a strong htopoI response that correlates with the onset of apoptotic hallmarks. Consequently, these results suggest that htopoI cleavage complex formation may be caused by signaling pathways independent of the kind of cellular stress. Thus, protein interactions or signaling cascades induced by DNA damage or cellular stress might lead to the formation of stabilized cleavage complexes rather than the DNA lesion itself. Finally, we show that p53 not only plays a key role in the regulation of the htopoI response to UV-C irradiation but also to treatment with colcemid.

The human topoisomerase I damage response plays a role in apoptosis

Previous studies have shown that human topoisomerase I cleavage complexes form as a response to various DNA damages in vivo, the so called human topoisomerase I “damage response”. It was suggested that this damage response may play a role in DNA repair as well as in apoptosis, but only very few investigations have been done and the significance of the damage response still remains unclear. Here we demonstrate that human topoisomerase I cleavage complexes induced by high doses of UV irradiation are highly stable for up to 48 h. Furthermore, we show that human topoisomerase I cleavage complexes correlate with apoptosis. However, at low UV doses the cleavage complex level was very low and the complexes were repaired. Surprisingly, we found that high levels of stable cleavage complexes were not only found in UV-irradiated cells but also in untreated cells that underwent apoptosis. A possible role of human topoisomerase I in apoptosis is discussed.

Scepticism and trust: two counterpoint essentials in science education for complex socio-scientific issues

In this response to Tom G. K. Bryce and Stephen P. Day’s (Cult Stud Sci Educ. doi:10.1007/s11422-013-9500-0, 2013) original article, I share with them their interest in the teaching of climate change in school science, but I widen it to include other contemporary complex socio-scientific issues that also need to be discussed. I use an alternative view of the relationship between science, technology and society, supported by evidence from both science and society, to suggest science-informed citizens as a more realistic outcome image of school science than the authors’ one of mini-scientists. The intellectual independence of students Bryce and Day assume, and intend for school science, is countered with an active intellectual dependence. It is only in relation to emerging and uncertain scientific contexts that students should be taught about scepticism, but they also need to learn when, and why to trust science as an antidote to the expressions of doubting it. Some suggestions for pedagogies that could lead to these new learnings are made. The very recent fifth report of the IPCC answers many of their concerns about climate change.

Mathematical investigation of the interactions between the inflammatory response and mechanical aspects of dermal wound repair

Dermal wound repair involves complex interactions between cells, cytokines and mechanics to close injuries to the skin. In particular, we investigate the contribution of fibroblasts, myofibroblasts, TGFβ, collagen and local tissue mechanics to wound repair in the human dermis. We develop a morphoelastic model where a realistic representation of tissue mechanics is key, and a fibrocontractive model that involves a reasonable approximation to the true kinetics of the important bioactive species. We use each of these descriptions to elucidate the mechanisms that generate pathologies such as hypertrophic scars, contractures and keloids. We find that for hypertrophic scar and contracture development, factors regulating the myofibroblast phenotype are critical, with heightened myofibroblast activation, reduced myofibroblast apoptosis or prolonged inflammation all predicted as mediators for scar hypertrophy and contractures. Prevention of these pathologies is predicted when myofibroblast apoptosis is induced, myofibroblast activation is blocked or TGFβ is neutralised. To investigate keloid invasion, we develop a caricature representation of the fibrocontractive model and find that TGFβ spread is the driving factor behind keloid growth. Blocking activation of TGFβ is found to cause keloid regression. Thus, we recommend myofibroblasts and TGFβ as targets for clinicians when developing intervention strategies for prevention and cure of fibrotic scars.

Design of a traumatic injury simulator for assessing lower limb response to high loading rates

Current military conflicts are characterized by the use of the improvised explosive device. Improvements in personal protection, medical care, and evacuation logistics have resulted in increasing numbers of casualties surviving with complex musculoskeletal injuries, often leading to lifelong disability. Thus, there exists an urgent requirement to investigate the mechanism of extremity injury caused by these devices in order to develop mitigation strategies. In addition, the wounds of war are no longer restricted to the battlefield; similar injuries can be witnessed in civilian centers following a terrorist attack. Key to understanding such mechanisms of injury is the ability to deconstruct the complexities of an explosive event into a controlled, laboratory-based environment. In this article, a traumatic injury simulator, designed to recreate in the laboratory the impulse that is transferred to the lower extremity from an anti-vehicle explosion, is presented and characterized experimentally and numerically. Tests with instrumented cadaveric limbs were then conducted to assess the simulator’s ability to interact with the human in two mounting conditions, simulating typical seated and standing vehicle passengers. This experimental device will now allow us to (a) gain comprehensive understanding of the load-transfer mechanisms through the lower limb, (b) characterize the dissipating capacity of mitigation technologies, and (c) assess the bio-fidelity of surrogates.

Complex regulation of membrane-type matrix metalloproteinase expression and matrix metalloproteinase-2 activation by concanavalin A in MDA-MB-231 human breast cancer cells

Matrix Metalloproteinase-2 (MMP-2) is secreted as a zymogen, the activation of which has been associated with metastatic progression in human breast cancer (HBC). Concanavalin A (Con A) has been found to induce activation of MMP-2 in invasive HBC cell lines. Con A effects on the expression of mRNA for membrane-type matrix metalloproteinase (MT-MMP), a newly described cell surface-associated MMP, showed a close temporal correlation with induction of MMP-2 activation. It is surprising that MT-MMP mRNA is constitutively present in the uninduced MDA-MB-231 cell, despite a lack of MMP-2 activation. We have used actinomycin D to demonstrate a partial requirement for de novo gene expression in the induction of MMP-2 activation by Con A in MDA-MB-231 HBC cells. Furthermore, this transcriptional response to Con A appeared to require the continued presence of Con A for its manifestation. The nontranscriptional component of the Con A induction manifests rapidly, is quite substantial, and persists strongly despite actinomycin D abrogation of both constitutive and Con A-induced MT-MMP. Cycloheximide analyses suggest that protein synthesis may be involved in this rapid transcription-independent response. These studies suggest that Con A induces MMP-2-activation in part by up-regulation of MT-MMP expression but has a more complicated mode of action, involving additional nontranscriptional effects, which apparently require protein synthesis.

Behavior of the electron temperature in nonuniform complex plasmas

The response of complex ionized gas systems to the presence of nonuniform distribution of charged grains is investigated using a kinetic model. Contrary to an existing view that the electron temperature inevitably increases in the grain-occupied region because of enhanced ionization to compensate for the electrons lost to the grains, it is shown that this happens only when the ionizing electric field increases in the electron depleted region. The results for two typical plasma systems suggest that when the ionizing electric field depends on the spatially averaged electron density, the electron temperature in the grain containing region can actually decrease.

Innate transcriptional networks activated in bladder in response to uropathogenic Escherichia coli drive diverse biological pathways and rapid synthesis of IL-10 for defense against bacterial urinary tract infection

Early transcriptional activation events that occur in bladder immediately following bacterial urinary tract infection (UTI) are not well defined. In this study, we describe the whole bladder transcriptome of uropathogenic Escherichia coli (UPEC) cystitis in mice using genome-wide expression profiling to define the transcriptome of innate immune activation stemming from UPEC colonization of the bladder. Bladder RNA from female C57BL/6 mice, analyzed using 1.0 ST-Affymetrix microarrays, revealed extensive activation of diverse sets of innate immune response genes, including those that encode multiple IL-family members, receptors, metabolic regulators, MAPK activators, and lymphocyte signaling molecules. These were among 1564 genes differentially regulated at 2 h postinfection, highlighting a rapid and broad innate immune response to bladder colonization. Integrative systems-level analyses using InnateDB (http://www.innatedb.com) bioinformatics and ingenuity pathway analysis identified multiple distinct biological pathways in the bladder transcriptome with extensive involvement of lymphocyte signaling, cell cycle alterations, cytoskeletal, and metabolic changes. A key regulator of IL activity identified in the transcriptome was IL-10, which was analyzed functionally to reveal marked exacerbation of cystitis in IL-10–deficient mice. Studies of clinical UTI revealed significantly elevated urinary IL-10 in patients with UPEC cystitis, indicating a role for IL-10 in the innate response to human UTI. The whole bladder transcriptome presented in this work provides new insight into the diversity of innate factors that determine UTI on a genome-wide scale and will be valuable for further data mining. Identification of protective roles for other elements in the transcriptome will provide critical new insight into the complex cascade of events that underpin UTI.

Primary and secondary neural networks of auditory prepulse inhibition: a functional magnetic resonance imaging study of sensorimotor gating of the human acoustic startle response

Feedforward inhibition deficits have been consistently demonstrated in a range of neuropsychiatric conditions using prepulse inhibition (PPI) of the acoustic startle eye-blink reflex when assessing sensorimotor gating. While PPI can be recorded in acutely decerebrated rats, behavioural, pharmacological and psychophysiological studies suggest the involvement of a complex neural network extending from brainstem nuclei to higher order cortical areas. The current functional magnetic resonance imaging study investigated the neural network underlying PPI and its association with electromyographically (EMG) recorded PPI of the acoustic startle eye-blink reflex in 16 healthy volunteers. A sparse imaging design was employed to model signal changes in blood oxygenation level-dependent (BOLD) responses to acoustic startle probes that were preceded by a prepulse at 120 ms or 480 ms stimulus onset asynchrony or without prepulse. Sensorimotor gating was EMG confirmed for the 120-ms prepulse condition, while startle responses in the 480-ms prepulse condition did not differ from startle alone. Multiple regression analysis of BOLD contrasts identified activation in pons, thalamus, caudate nuclei, left angular gyrus and bilaterally in anterior cingulate, associated with EMGrecorded sensorimotor gating. Planned contrasts confirmed increased pons activation for startle alone vs 120-ms prepulse condition, while increased anterior superior frontal gyrus activation was confirmed for the reverse contrast. Our findings are consistent with a primary pontine circuitry of sensorimotor gating that interconnects with inferior parietal, superior temporal, frontal and prefrontal cortices via thalamus and striatum. PPI processes in the prefrontal, frontal and superior temporal cortex were functionally distinct from sensorimotor gating.

Improving the police response to online fraud

Online fraud poses a significant problem to society in terms of its monetary losses and the devastating impact on victims. It also poses significant challenges to law enforcement agencies, regarding their ability to investigate crimes which are complex, occur in a virtual environment, incorporate multiple (often international) jurisdictions, and have a very low reporting rate. This paper examines the police response to online fraud. It argues that traditionally, fraud has received little attention and priority from police agencies and this is exacerbated in the online context. In contrast to this, the paper presents the example of Project Sunbird, a partnership between the West Australian Police and the West Australian Department of Commerce which has embraced the use of financial intelligence to proactively contact suspected victims of online fraud. This paper argues that a proactive approach to policing online fraud can have substantial positive effects for police and victims alike.

Identification of a BRCA1-mRNA splicing complex required for efficient DNA repair and maintenance of genomic stability

Mutations within BRCA1 predispose carriers to a high risk of breast and ovarian cancers. BRCA1 functions to maintain genomic stability through the assembly of multiple protein complexes involved in DNA repair, cell-cycle arrest, and transcriptional regulation. Here, we report the identification of a DNA damage-induced BRCA1 protein complex containing BCLAF1 and other key components of the mRNA-splicing machinery. In response to DNA damage, this complex regulates pre-mRNA splicing of a number of genes involved in DNA damage signaling and repair, thereby promoting the stability of these transcripts/proteins. Further, we show that abrogation of this complex results in sensitivity to DNA damage, defective DNA repair, and genomic instability. Interestingly, mutations in a number of proteins found within this complex have been identified in numerous cancer types. These data suggest that regulation of splicing by the BRCA1-mRNA splicing complex plays an important role in the cellular response to DNA damage.

Energy gap in the aetiology of body weight gain and obesity: A challenging concept with a complex evaluation and pitfalls

The concept of energy gap(s) is useful for understanding the consequence of a small daily, weekly, or monthly positive energy balance and the inconspicuous shift in weight gain ultimately leading to overweight and obesity. Energy gap is a dynamic concept: an initial positive energy gap incurred via an increase in energy intake (or a decrease in physical activity) is not constant, may fade out with time if the initial conditions are maintained, and depends on the 'efficiency' with which the readjustment of the energy imbalance gap occurs with time. The metabolic response to an energy imbalance gap and the magnitude of the energy gap(s) can be estimated by at least two methods, i.e. i) assessment by longitudinal overfeeding studies, imposing (by design) an initial positive energy imbalance gap; ii) retrospective assessment based on epidemiological surveys, whereby the accumulated endogenous energy storage per unit of time is calculated from the change in body weight and body composition. In order to illustrate the difficulty of accurately assessing an energy gap we have used, as an illustrative example, a recent epidemiological study which tracked changes in total energy intake (estimated by gross food availability) and body weight over 3 decades in the US, combined with total energy expenditure prediction from body weight using doubly labelled water data. At the population level, the study attempted to assess the cause of the energy gap purported to be entirely due to increased food intake. Based on an estimate of change in energy intake judged to be more reliable (i.e. in the same study population) and together with calculations of simple energetic indices, our analysis suggests that conclusions about the fundamental causes of obesity development in a population (excess intake vs. low physical activity or both) is clouded by a high level of uncertainty.

Effects of particle size distribution in the response of model granular materials in multi-ring shear

Rail track undergoes complex loading patterns under moving traffic conditions compared to roads due to its continued and discontinued multi-layered structure, including rail, sleepers, ballast layer, sub-ballast layer, and subgrade. Particle size distributions (PSDs) of ballast, subballast, and subgrade layers can be critical in cyclic plastic deformation of rail track under moving traffic on frequent track degradation of rail tracks, especially at bridge transition zones. Conventional test approaches: static shear and cyclic single-point load tests are however unable to replicate actual loading patterns of moving train. Multi-ring shear apparatus; a new type of torsional simple shear apparatus, which can reproduce moving traffic conditions, was used in this study to investigate influence of particle size distribution of rail track layers on cyclic plastic deformation. Three particle size distributions, using glass beads were examined under different loading patterns: cyclic sin-gle-point load, and cyclic moving wheel load to evaluate cyclic plastic deformation of rail track under different loading methods. The results of these tests suggest that particle size distributions of rail track structural layers have significant impacts on cyclic plastic deformation under moving train load. Further, the limitations in con-ventional test methods used in laboratories to estimate the plastic deformation of rail track materials lead to underestimate the plastic deformation of rail tracks.

Anticipatory and reactive response to falls: Muscle synergy activation of forearm muscles

We investigated the surface electromyogram response of six forearm muscles to falls onto the outstretched hand. The extensor carpi radialis longus, extensor carpi radialis brevis, extensor carpi ulnaris, abductor pollicis longus, flexor carpi radialis and flexor carpi ulnaris muscles were sampled from eight volunteers who underwent ten self-initiated falls. All muscles initiated prior to impact. Co-contraction is the most obvious surface electromyogram feature. The predominant response is in the radial deviators. The surface electromyogram timing we recorded would appear to be a complex anticipatory response to falling modified by the ef- fect on the forearm muscles following impact. The mitigation of the force of impact is probably more importantly through shoulder abduction and extension and elbow flexion rather than action of the forearm muscles.

Mandatory Reporting and the Difficulties Identifying and Responding to Risk of Severe Neglect: A Response Requiring a Rethink

Neglect of children is a significant social issue worldwide and is typically the most frequently reported form of maltreatment in Western nations, with its severe forms sometimes resulting in significant illness and disablement or death. Yet, paradoxically, it remains ‘neglected’ and largely in the shadow of physical and sexual abuse, often being viewed as less serious despite the real-life consequences of its insidious and compounding nature and the lasting damage it causes to intergenerational familial relationships and the life outcomes of those affected. This chapter explores the many complex forms of child neglect, its causes and impacts and the strategies to prevent it. In particular, a critical standpoint is taken in analysing the rationale and merits of mandatory reporting of neglect and their effects, systemically and for children.