USP50 suppresses alternative RecQ helicase use and deleterious DNA2 activity during replication.

Mammalian DNA replication employs several RecQ DNA helicases to orchestrate the faithful duplication of genetic information. Helicase function is often coupled to the activity of specific nucleases, but how helicase and nuclease activities are co-directed is unclear. Here we identify the inactive ubiquitin-specific protease, USP50, as a ubiquitin-binding and chromatin-associated protein required for ongoing replication, fork restart, telomere maintenance and cellular survival during replicative stress. USP50 supports WRN:FEN1 at stalled replication forks, suppresses MUS81-dependent fork collapse and restricts double-strand DNA breaks at GC-rich sequences. Surprisingly we find that cells depleted for USP50 and recovering from a replication block exhibit increased DNA2 and RECQL4 foci and that the defects in ongoing replication, poor fork restart and increased fork collapse seen in these cells are mediated by DNA2, RECQL4 and RECQL5. These data define a novel ubiquitin-dependent pathway that promotes the balance of helicase: nuclease use at ongoing and stalled replication forks.

Mutant p53 promotes RCP-dependent chemoresistance coinciding with increased delivery of P-glycoprotein to the plasma membrane.

TP53 is the most frequently mutated gene in cancers. Mutations lead to loss of p53 expression or expression of a mutant protein. Mutant p53 proteins commonly lose wild-type function, but can also acquire novel functions in promoting metastasis and chemoresistance. Previously, we uncovered a role for Rab-coupling protein (RCP) in mutant p53-dependent invasion. RCP promotes endosomal recycling and signalling of integrins and receptor tyrosine kinases. In a screen to identify novel RCP-interacting proteins, we discovered P-glycoprotein (P-gp). Thus, we hypothesised that mutant p53 could promote chemoresistance through RCP-dependent recycling of P-gp. The interaction between RCP and P-gp was verified endogenously and loss of RCP or mutant p53 rendered cells more sensitive to cisplatin and etoposide. In mutant p53 cells we detected an RCP-dependent delivery of P-gp to the plasma membrane upon drug treatment and decreased retention of P-gp substrates. A co-localisation of P-gp and RCP was seen in mutant p53 cells, but not in p53-null cells upon chemotherapeutic exposure. In conclusion, mutant p53 expression enhanced co-localisation of P-gp and RCP to allow for rapid delivery of P-gp to the plasma membrane and increased resistance to chemotherapeutics.

Combined bezafibrate, medroxyprogesterone acetate and valproic acid treatment inhibits osteosarcoma cell growth without adversely affecting normal mesenchymal stem cells.

Drug repurposing is a cost-effective means of targeting new therapies for cancer. We have examined the effects of the repurposed drugs, bezafibrate, medroxyprogesterone acetate and valproic acid on human osteosarcoma cells, i.e., SAOS2 and MG63 compared with their normal cell counterparts, i.e. mesenchymal stem/stromal cells (MSCs). Cell growth, viability and migration were measured by biochemical assay and live cell imaging, whilst levels of lipid-synthesising enzymes were measured by immunoblotting cell extracts. These drug treatments inhibited the growth and survival of SAOS2 and MG63 cells most effectively when used in combination (termed V-BAP). In contrast, V-BAP treated MSCs remained viable with only moderately reduced cell proliferation. V-BAP treatment also inhibited migratory cell phenotypes. MG63 and SAOS2 cells expressed much greater levels of fatty acid synthase and stearoyl CoA desaturase 1 than MSCs, but these elevated enzyme levels significantly decreased in the V-BAP treated osteosarcoma cells prior to cell death. Hence, we have identified a repurposed drug combination that selectively inhibits the growth and survival of human osteosarcoma cells in association with altered lipid metabolism without adversely affecting their non-transformed cell counterparts.

Does intensive multidisciplinary intervention for adults who elect bariatric surgery improve post-operative weight loss, co-morbidities, and quality of life? A systematic review and meta-analysis.

This systematic review and meta-analysis of intervention studies aims to evaluate the effect of preoperative and/or post-operative support for adults who elect bariatric surgery delivered by a multidisciplinary team (MDT) on post-operative body composition, mental health, co-morbidities, quality of life, and side effects. Six electronic databases were searched. Revman and GRADE were used to assess confidence in pooled effects. Included interventions (N = 1533 participants in total) focused on lifestyle counselling (n = 4 studies), psychology (n = 4 studies), or exercise (n = 10 studies); comparator groups were less intensive usual care. Intensive MDT interventions increased post-operative weight loss (SMD: -0.94; 95% CI: -1.27 to -0.61) if delivered post-operatively. Preoperative and post-operative intensive interventions improved symptoms of depression and anxiety, quality of life, diastolic blood pressure, and resting heart rate but not lipids or glycaemic measures. Whilst usual MDT care is important preoperatively, this review conditionally recommends intensive MDT interventions for enhanced post-operative weight loss if delivered in the post-operative period, led by any health professional, based on moderate evidence. This review also conditionally recommends preoperative and/or post-operative lifestyle, nutrition, or psychology counselling and/or physical activity for improved mental and physical health. Further randomized controlled trials are required, which aim to specifically evaluate the best use of MDT resources.

Isomerization of BRCA1-BARD1 promotes replication fork protection.

The integrity of genomes is constantly threatened by problems encountered by the replication fork. BRCA1, BRCA2 and a subset of Fanconi anaemia proteins protect stalled replication forks from degradation by nucleases, through pathways that involve RAD51. The contribution and regulation of BRCA1 in replication fork protection, and how this role relates to its role in homologous recombination, is unclear. Here we show that BRCA1 in complex with BARD1, and not the canonical BRCA1-PALB2 interaction, is required for fork protection. BRCA1-BARD1 is regulated by a conformational change mediated by the phosphorylation-directed prolyl isomerase PIN1. PIN1 activity enhances BRCA1-BARD1 interaction with RAD51, thereby increasing the presence of RAD51 at stalled replication structures. We identify genetic variants of BRCA1-BARD1 in patients with cancer that exhibit poor protection of nascent strands but retain homologous recombination proficiency, thus defining domains of BRCA1-BARD1 that are required for fork protection and associated with cancer development. Together, these findings reveal a BRCA1-mediated pathway that governs replication fork protection.

Establishing an evidenced-based dietetic model of care in haemodialysis using implementation science.

AIM: To establish an evidence-based dietetics service in an in-centre haemodialysis unit utilising implementation science. METHODS: The service was developed through the Knowledge-to-Action Framework. The steps of the Action Cycle were addressed through a literature review, identification of evidence-based guidelines, benchmarking and local staff engagement. The theoretical domains framework (TDF) was used to identify barriers/enablers, and behaviour change wheel to determine appropriate interventions. To monitor, evaluate outcomes and assess sustained knowledge use we employed multidisciplinary team engagement and database use. Audit data were collected at baseline, 6 and 12 months on nutrition assessment (Patient-Generated Subjective Global Assessment), intervention timeliness and alignment to dietetic workforce recommendations. Descriptive statistics, McNemar tests and a linear mixed model were applied. RESULTS: Barriers existed in the knowledge, skills, environmental context and resources TDF domains. Suitable interventions were identified with training on nutritional management of haemodialysis patients delivered to 148 nurses, and nutrition management recommendations summarised into local procedural resources. A database to prompt and monitor outcome measures was created and indicated that over 18 months post-service commencement, eligible patients received nutrition assessment at least 6-monthly, aligning with recommendations. Prevalence of malnutrition was 28% (n = 9/32) at baseline, 23% (n = 5/22) at 6 months and 20% (n = 4/20) at 12 months (P = 0.50). CONCLUSIONS: We demonstrated benefits to service development and implementation with implementation science providing a structured and methodical approach to translating guidelines into practice. Development of training, resources and prompts for outcome measures has supported the establishment of an evidence-based dietetics service in a haemodialysis unit.

Biological relevance of cell-in-cell in cancers.

Cell-in-cell (CIC) is a term used to describe the presence of one, usually living, cell inside another cell that is typically considered non-phagocytic. Examples of this include tumour cells inside tumour cells (homotypic), mesenchymal stem cells inside tumour cells (heterotypic) or immune cells inside tumour cells (heterotypic). CIC formation can occur in cell lines and in tissues and it has been most frequently observed during inflammation and in cancers. Over the past 10 years, many researchers have studied CIC structures and a few different models have been proposed through which they can be formed, including entosis, cannibalism and emperipolesis among others. Recently, our laboratory discovered a role for mutant p53 in facilitating the formation of CIC and promoting genomic instability. These data and research by many others have uncovered a variety of molecules involved in CIC formation and have started to give us an idea of why they are formed and how they could contribute to oncogenic processes. In this perspective, we summarise current literature and speculate on the role of CIC in cancer biology.

The deSUMOylase SENP2 coordinates homologous recombination and nonhomologous end joining by independent mechanisms.

SUMOylation (small ubiquitin-like modifier) in the DNA double-strand break (DSB) response regulates recruitment, activity, and clearance of repair factors. However, our understanding of a role for deSUMOylation in this process is limited. Here we identify different mechanistic roles for deSUMOylation in homologous recombination (HR) and nonhomologous end joining (NHEJ) through the investigation of the deSUMOylase SENP2. We found that regulated deSUMOylation of MDC1 prevents excessive SUMOylation and its RNF4-VCP mediated clearance from DSBs, thereby promoting NHEJ. In contrast, we show that HR is differentially sensitive to SUMO availability and SENP2 activity is needed to provide SUMO. SENP2 is amplified as part of the chromosome 3q amplification in many cancers. Increased SENP2 expression prolongs MDC1 focus retention and increases NHEJ and radioresistance. Collectively, our data reveal that deSUMOylation differentially primes cells for responding to DSBs and demonstrates the ability of SENP2 to tune DSB repair responses.

A systematic review, meta-analysis and meta-regression of the prevalence of protein-energy malnutrition: associations with geographical region and sex.

Background: protein-energy malnutrition is a major health concern in home-dwelling older adults, particularly in the context of an ageing population. Therefore, a systematic review and meta-analysis was undertaken to determine the prevalence of malnutrition among older adults living independently in the community according to geographical region, sampling frame, rurality and sex. Methods: six electronic databases were searched until September 2016. Original research studies which used the Mini Nutritional Assessment, Patient-Generated Subjective Global Assessment or Subjective Global Assessment to determine nutrition status in community samples with a mean age of ≥65 years were critically appraised and pooled using meta-analysis. Meta-regression was used to explore predictors of malnutrition prevalence in pooled statistics with high heterogeneity. Results: 111 studies from 38 countries (n = 69,702 participants) were included. The pooled prevalence of malnutrition in the older community setting ranged from 0.8% (95% CI: 0.2-1.7%) in Northern Europe to 24.6% (95% CI: 0.0-67.9%) in South-East Asia. Of all sampling frames, participants receiving homecare services had the highest prevalence at 14.6% (95% CI: 9.9-20.0%). Malnutrition prevalence in rural communities (9.9%; 95% CI: 4.5-16.8%) was double that in urban communities (5.7%; 95% CI: 4.2-7.3%) and higher among females than males (odds ratio = 1.45 [95% CI: 1.27-1.66]; P < 0.00001). Conclusions: the results of this review provide strategic insight to develop public and community health priorities for preventing malnutrition and associated poor health outcomes.

Genomic instability in mutant p53 cancer cells upon entotic engulfment.

Cell-in-cell (CIC) structures are commonly seen in tumours. Their biological significance remains unclear, although they have been associated with more aggressive tumours. Here we report that mutant p53 promotes CIC via live cell engulfment. Engulfed cells physically interfere in cell divisions of host cells and for cells without p53 this leads to host cell death. In contrast, mutant p53 host cells survive, display aberrant divisions, multinucleation and tripolar mitoses. In xenograft studies, CIC-rich p53 mutant/null co-cultures show enhanced tumour growth. Furthermore, our results show that CIC is common within lung adenocarcinomas, is an independent predictor of poor outcome and disease recurrence, is associated with mutant p53 expression and correlated to measures of heterogeneity and genomic instability. These findings suggest that pro-tumorigenic entotic engulfment activity is associated with mutant p53 expression, and the two combined are a key factor in genomic instability.

Publisher Correction: Genomic instability in mutant p53 cancer cells upon entotic engulfment.

The original version of this article incorrectly omitted an affiliation of Patricia A. J. Muller: 'Cancer Research UK Manchester Institute, The University of Manchester | Alderley Park, Manchester, SK10 4TG, UK'. This has been corrected in both the PDF and HTML versions of the Article.

Is telehealth effective in managing malnutrition in community-dwelling older adults? A systematic review and meta-analysis.

Telehealth offers a feasible method to provide nutrition support to malnourished older adults. This systematic review and meta-analysis aims to determine the efficacy of telehealth methods in delivering malnutrition-related interventions to community-dwelling older adults. Studies in any language were searched in five electronic databases from inception to 2nd November 2017. Quality of the evidence was assessed using the Cochrane Risk of Bias tool and the GRADE approach. Nine studies were identified, with results published across 13 included publications, which had mostly low to unclear risk of bias. There were two interventions delivered to disease-specific groups, one with kidney disease and one with cancer; the remaining seven interventions were delivered to patients with mixed morbidities following discharge from an inpatient facility. Seven studies delivered telehealth via telephone consultations and two used internet-enabled telemedicine devices. Ten meta-analyses were performed. Malnutrition-focused telehealth interventions were found to improve protein intake in older adults by 0.13 g/kg body weight per day ([95%CI: 0.01-0.25]; P = .03; n = 2 studies; n = 200 participants; I2 = 41%; GRADE level: low) and to improve quality of life (standardised mean difference: 0.55 [95%CI: 0.11-0.99]; P = .01; n = 4 studies with n = 9 quality-of-life tools; n = 248 participants; I2 = 84%: GRADE level: very low). There were also trends towards improved nutrition status, physical function, energy intake, hospital readmission rates and mortality in the intervention groups. Overall, this review found telehealth is an effective method to deliver malnutrition-related interventions to older adults living at home, and is likely to result in clinical improvements compared with usual care or no intervention. However, further research with larger samples and stronger study designs are required to strengthen the body of evidence.

The effects of shiatsu: findings from a two-country exploratory study.

OBJECTIVES: To provide insight into client and practitioner perceptions of the effects of shiatsu, in the short and longer term, and positive and negative in nature. DESIGN: A two-country, exploratory study was undertaken in the United Kingdom and Germany. In-depth interviews were undertaken with a purposive sample of 14 shiatsu practitioners and 15 clients. Client interviews focused on the experience of shiatsu and perceptions of its effects, both positive and negative. Practitioners were also asked about factors that enhanced or inhibited successful treatment. The taped and transcribed data were analyzed using grounded theory, assisted by NVivo (QSR, Markham, Ontario, Canada) software. To enhance generalizability, the findings from the alternative country data set were presented to a further set of practitioners in each country and as a whole to an international meeting of practitioners from seven European countries. RESULTS: There was similarity in the perspectives of the clients and practitioners and participants from the United Kingdom and Germany. Both described a wide range of common, immediate and longer term effects. These included effects on initial symptoms, relaxation, sleeping, posture, and experiences of the body. A category of transitional effect arose, describing an effect that was not particularly positive and did not last long. Practitioners characterized this as being part of the healing response. Only a few negative effects were described by clients. One mentioned a negative physical reaction and two indicated difficulties coping with emotional reactions. While most practitioners conceived negative effects to be possible, these were more likely to be described as negative reactions. CONCLUSION: This exploratory study has shed greater light on the effects of shiatsu. The sample findings provide a user and practitioner grounded base for the design of appropriate questions for exploration in a larger and more generalizable study of the effects of shiatsu.

Appraisal of published reviews of research on psychotherapy and counseling with adults 1990-1998.

Reviews and meta-analyses of research on adult psychotherapy and counseling with adults published in English from 1990 to 1998 (N = 255) were rated according to the criteria advanced by A. D. Oxman and G. H. Guyatt (1988). A majority of the reviews fell substantially short of these basic standards; on average, review articles met only half of the 8 criteria, and only 20% met 7 or more of the criteria. Common deficiencies included failure to describe comprehensive search methods used to locate studies; failure to explain inclusion and exclusion criteria for studies; and failure to provide reliable, unbiased assessments of the studies reviewed. The reviews showed no tendency to improve over the period studied.