Polo-like kinase 3 regulates CtIP during DNA double-strand break repair in G1

DNA double-strand breaks (DSBs) are repaired by nonhomologous end joining (NHEJ) or homologous recombination (HR). The C terminal binding protein–interacting protein (CtIP) is phosphorylated in G2 by cyclin-dependent kinases to initiate resection and promote HR. CtIP also exerts functions during NHEJ, although the mechanism phosphorylating CtIP in G1 is unknown. In this paper, we identify Plk3 (Polo-like kinase 3) as a novel DSB response factor that phosphorylates CtIP in G1 in a damage-inducible manner and impacts on various cellular processes in G1. First, Plk3 and CtIP enhance the formation of ionizing radiation-induced translocations; second, they promote large-scale genomic deletions from restriction enzyme-induced DSBs; third, they are required for resection and repair of complex DSBs; and finally, they regulate alternative NHEJ processes in Ku−/− mutants. We show that mutating CtIP at S327 or T847 to nonphosphorylatable alanine phenocopies Plk3 or CtIP loss. Plk3 binds to CtIP phosphorylated at S327 via its Polo box domains, which is necessary for robust damage-induced CtIP phosphorylation at S327 and subsequent CtIP phosphorylation at T847

Alternate wetting and drying irrigation maintained rice yields despite half the irrigation volume, but is currently unlikely to be adopted by smallholder lowland rice farmers in Nepal

Alternate wetting and drying (AWD) irrigation can save water while maintaining rice yields, but in some countries its adoption by farmers remains limited. Key knowledge gaps include the effect of AWD on early vegetative vigor and its relationship with yield; the effects of AWD on yield and water use efficiency of local cultivars used by smallholder farmers; and the socio-economic factors influencing current irrigation scheduling. To address these questions, an on-farm field trial of dry-season (chaite) rice, comparing two locally important cultivars (Hardinath-1 and CH-45) under AWD imposed from 1 week after transplanting to flowering and continuous flooding (CF), was carried out in Agyauli in the central Terai region of Nepal, and triangulated with social research methods exploring the rationale for current irrigation scheduling and perceptions of AWD. Although AWD plots received on average 57% less irrigation water than CF plots, yields did not significantly differ between irrigation treatments, indicating that AWD could considerably enhance crop water use efficiency in this region. In the earlier flowering, more vigorous CH-45, there were no treatment differences in any yield component while in the later flowering Hardinath-1, an 11% decrease in filled grain number was compensated by a 14% increase in the percentage of effective tillers per hill. Although leaf elongation rate on the main tiller did not differ between treatments, tillering and green fraction (a measure of canopy closure) were significantly higher under AWD. Surveys established that most local farmers are already using a local adaptation of AWD to modify irrigation volumes, in some cases in response to a limited and unreliable water supply. However, farmers have few direct incentives to reduce overall water use under current water governance, and formal AWD practices are therefore unlikely to be adopted despite their viability as a water-saving irrigation technique

Breathlessness and inflammation: relationship and implications

Purpose of Review: Breathlessness and chronic inflammation both span a wide range of disease contexts and hold prognostic significance. The possibility of a causal relationship between the two has been hypothesised. The aims of this article are 1) to review the intersections between breathlessness and inflammation in the literature, 2) to describe potential mechanisms connecting the two phenomena, and 3) to discuss the potential clinical implications of a causal relationship. Recent findings: There is a very limited literature exploring the relationship between systemic inflammation and breathlessness in COPD, heart failure and cancer. One large study in cancer patients is suggestive of a weak association between self-reported breathlessness and inflammation. Studies exploring the relationship between inflammation and MRC Dyspnoea grade have produced inconsistent findings. Though a causal relationship has not yet been demonstrated, this relationship might be mediated through the effects of both inflammation and breathlessness on the skeletal muscle and stress hormone systems. Summary: There is much progress to be made in this area. Interventional studies, evaluating the impact of anti-inflammatory interventions on breathlessness, are needed to help determine whether a causal relationship exists. If proven, this relationship might have important implications for both the treatment and impact of breathlessness

DNA double-strand break resection occurs during non-homologous end joining in G1 but is distinct from resection during homologous recombination

Canonical non-homologous end joining (c-NHEJ) repairs DNA double-strand breaks (DSBs) in G1 cells with biphasic kinetics. We show that DSBs repaired with slow kinetics, including those localizing to heterochromatic regions or harboring additional lesions at the DSB site, undergo resection prior to repair by c-NHEJ and not alt-NHEJ. Resection-dependent c-NHEJ represents an inducible process during which Plk3 phosphorylates CtIP, mediating its interaction with Brca1 and promoting the initiation of resection. Mre11 exonuclease, EXD2, and Exo1 execute resection, and Artemis endonuclease functions to complete the process. If resection does not commence, then repair can ensue by c-NHEJ, but when executed, Artemis is essential to complete resection-dependent c-NHEJ. Additionally, Mre11 endonuclease activity is dispensable for resection in G1. Thus, resection in G1 differs from the process in G2 that leads to homologous recombination. Resection-dependent c-NHEJ significantly contributes to the formation of deletions and translocations in G1, which represent important initiating events in carcinogenesis

Advancing the application of systems thinking in health: managing rural China health system development in complex and dynamic contexts

Background: This paper explores the evolution of schemes for rural finance in China as a case study of the long and complex process of health system development. It argues that the evolution of these schemes has been the outcome of the response of a large number of agents to a rapidly changing context and of efforts by the government to influence this adaptation process and achieve public health goals. Methods:The study draws on several sources of data including a review of official policy documents and academic papers and in-depth interviews with key policy actors at national level and at a sample of localities. Results: The study identifies three major transition points associated with changes in broad development strategy and demonstrates how the adaptation of large numbers of actors to these contextual changes had a major impact on the performance of the health system. Further, it documents how the Ministry of Health viewed its role as both an advocate for the interests of health facilities and health workers and as the agency responsible for ensuring that government health system objectives were met. It is argued that a major reason for the resilience of the health system and its ability to adapt to rapid economic and institutional change was the ability of the Ministry to provide overall strategy leadership. Additionally, it postulates that a number of interest groups have emerged, which now also seek to influence the pathway of health system development. Conclusions: This history illustrates the complex and political nature of the management of health system development and reform. The paper concludes that governments will need to increase their capacity to analyze the health sector as a complex system and to manage change processes.UKaid: DFI

Addressing Antimicrobial Resistance in China: Policy Implementation in a Complex Context

The effectiveness of antibiotics in treating bacterial infections is decreasing in China because of the widespread development of resistant organisms. Although China has enacted a number of regulations to address this problem, but the impact is very limited. This paper investigates the implementation of these regulations through the lens of complex adaptive systems (CAS). It presents the findings from reviews of relevant policy documents and published papers. The paper identifies different types of agent and explores their interaction with regard to the use of antibiotics and their responses to changes of the regulations. It focuses particularly on the impact of perverse financial incentives on overall patterns of use of antibiotics. Implications for the possibilities of nonlinear results, interactive relationships, and new pathways of policy implementation are discussed. The paper concludes that policy-makers need to better understand the objectives, incentives and potential adaptive behaviors of the agents when they implement interventions to improve antibiotic use and reduce the risk of emergence of resistant organism

Author response

Previously we observed differential activation in individual columns of the periaqueductal grey (PAG) during breathlessness and its conditioned anticipation (Faull et al., 2016). Here, we have extended this work by determining how the individual columns of the PAG interact with higher cortical centres, both at rest and in the context of breathlessness threat. Activation was observed in ventrolateral PAG (vlPAG) and lateral PAG (lPAG), where activity scaled with breathlessness intensity ratings, revealing a potential interface between sensation and cognition during breathlessness. At rest the lPAG was functionally correlated with cortical sensorimotor areas, conducive to facilitating fight/flight responses, and demonstrated increased synchronicity with the amygdala during breathlessness. The vlPAG showed fronto-limbic correlations at rest, whereas during breathlessness anticipation, reduced functional synchronicity was seen to both lPAG and motor structures, conducive to freezing behaviours. These results move us towards understanding how the PAG might be intricately involved in human responses to threat