Rac1 plays a role in CXCL12 but not CCL3-induced chemotaxis and Rac1 GEF inhibitor NSC23766 has off target effects on CXCR4

Cell mi­gra­tion to­wards a chemo­tac­tic stim­u­lus re­lies on the re-arrange­ment of the cy­toskele­ton, which is trig­gered by ac­ti­va­tion of small G pro­teins RhoA, Rac1 and Cd­c42, and leads to for­ma­tion of lamel­lopo­dia and actin poly­meri­sa­tion amongst other ef­fects. Here we show that Rac1 is im­por­tant for CX­CR4 in­duced chemo­taxis but not for CCR1/​CCR5 in­duced chemo­taxis. For CX­CL12-in­duced mi­gra­tion via CX­CR4, breast can­cer MCF-7 cells are re­liant on Rac1, sim­i­larly to THP-1 mono­cytes and Ju­rkat T-cells. For CCL3-in­duced mi­gra­tion via CCR1 and/​or CCR5, Rac1 sig­nalling does not reg­u­late cell mi­gra­tion in ei­ther sus­pen­sion or ad­her­ent cells. We have con­firmed the in­volve­ment of Rac1 with the use of a spe­cific Rac1 block­ing pep­tide. We also used a Rac1 in­hibitor EHT 1864 and a Rac1-GEF in­hibitor NSC23766 to probe the im­por­tance of Rac1 in chemo­taxis. Both in­hibitors did not block CCL3-in­duced chemo­taxis, but they were able to block CX­CL12-in­duced chemo­taxis. This con­firms that Rac1 ac­ti­va­tion is not es­sen­tial for CCL3-in­duced mi­gra­tion, how­ever NSC23766 might have sec­ondary ef­fects on CX­CR4. This small mol­e­cule ex­hibits ag­o­nis­tic fea­tures in in­ter­nal­i­sa­tion and cAMP as­says, whereas it acts as an an­tag­o­nist for CX­CR4 in mi­gra­tion and cal­cium re­lease as­says. Our find­ings strongly sug­gest that Rac1 ac­ti­va­tion is not nec­es­sary for CCL3 sig­nalling, and re­veal that NSC23766 could be a novel CX­CR4 re­cep­tor lig­and

Improving identification of cognitive impairment in fragility fracture patients : impact of educational guidelines on current practice

Peer reviewedPreprintPublisher PD

Statistical Aspects of the Use of Biomarkers in Nutritional Epidemiology Research

Few strong and consistent associations have arisen from observational studies of dietary consumption in relation to chronic disease risk. Measurement error in self-reported dietary assessment may be obscuring many such associations. Attempts to correct for measurement error have mostly used a second self-report assessment in a subset of a study cohort to calibrate the self-report assessment used throughout the cohort, under the dubious assumption of uncorrelated measurement errors between the two assessments. The use, instead, of objective biomarkers of nutrient consumption to produce calibrated consumption estimates provides a promising approach to enhance study reliability. As summarized here, we have recently applied this nutrient biomarker approach to examine energy, protein, and percent of energy from protein, in relation to disease incidence in Women’s Health Initiative cohorts, and find strong associations that are not evident without biomarker calibration. A major bottleneck for the broader use of a biomarker-calibration approach is the rather few nutrients for which a suitable biomarker has been developed. Some methodologic approaches to the development of additional pertinent biomarkers, including the possible use of a respiratory quotient from indirect calorimetry for macronutrient biomarker development, and the potential of human feeding studies for the evaluation of a range of urine- and blood-based potential biomarkers, will briefly be described

Blurring of boundaries in the doctor-patient relationship

No description supplie

Communicating the Oncotype DX® test results: a novel educational workshop for specialist breast care nurses

No description supplie

Cytotoxic activity of marine sponge extracts from the sub-Antarctic Islands and the Southern Ocean

publisher versionOver the past 50 years, marine invertebrates, especially sponges, have proven to be a valuable source of new and/or bioactive natural products that have the potential to be further developed as lead compounds for pharmaceutical applications. Although marine benthic invertebrate communities occurring off the coast of South Africa have been explored for their biomedicinal potential, the natural product investigation of marine sponges from the sub-Antarctic Islands in the Southern Ocean for the presence of bioactive secondary metabolites has been relatively unexplored thus far. We report here the results for the biological screening of both aqueous and organic extracts prepared from nine specimens of eight species of marine sponges, collected from around Marion Island and the Prince Edward Islands in the Southern Ocean, for their cytotoxic activity against three cancer cell lines. The results obtained through this multidisciplinary collaborative research effort by exclusively South African institutions has provided an exciting opportunity to discover cytotoxic compounds from sub-Antarctic sponges, whilst contributing to our understanding of the biodiversity and geographic distributions of these cold-water invertebrates. Therefore, we acknowledge here the various contributions of the diverse scientific disciplines that played a pivotal role in providing the necessary platform for the future natural products chemistry investigation of these marine sponges from the sub- Antarctic Islands and the Southern Ocean. Significance: This study will contribute to understanding the biodiversity and geographic distributions of sponges in the Southern Ocean. This multidisciplinary project has enabled the investigation of marine sponges for the presence of cytotoxic compounds. Further investigation will lead to the isolation and identification of cytotoxic compounds present in the active sponge extracts.University of Cape Town; South African Medical Research Council; National Research Foundation (South Africa); CANSA; Rhodes University; Department of Science and Technology; Department of Environmental Affairs; SANA

Clinical characteristics associated with the prescribing of SSRI medication in adolescents with major unipolar depression.

Unipolar major depressions (MD) emerge markedly during adolescence. National Institute for Health and Care Excellence (NICE) UK recommends psychological therapies, with accompanying selective serotonin reuptake inhibitors (SSRIs) prescribed in severe cases only. Here, we seek to determine the extent and rationale of SSRI prescribing in adolescent MD before entering a randomised clinical trial. SSRI prescribing, together with their clinical characteristics was determined in 465 adolescent patients with MD prior to receiving a standardised psychological therapy as part of the Improving mood with psychoanalytic and cognitive therapies (IMPACT) clinical trial. Overall, 88 (19 %) had been prescribed antidepressants prior to psychological treatment. The clinical correlates varied by gender: respectively, depression severity in boys and self-harming behaviours in girls. Prescribing also differed between clinical research centres. Medical practitioners consider severity of depression in boys as an indicator for antidepressant prescribing. Self-injury in girls appears to be utilised as a prescribing aid which is inconsistent with past and current revised UK NICE guidelines.RCT Study supported by a grant to IMG (Chief Investigator) from the NIHR-HTA (trial number ISRCTN83033550, grant number 06/05/01).This is the author accepted manuscript. It is currently under an indefinite embargo pending publication by Springer

Evaluation of polygenic risk scores for breast and ovarian cancer risk prediction in BRCA1 and BRCA2 mutation carriers

Background: Genome-wide association studies (GWAS) have identified 94 common single-nucleotide polymorphisms (SNPs) associated with breast cancer (BC) risk and 18 associated with ovarian cancer (OC) risk. Several of these are also associated with risk of BC or OC for women who carry a pathogenic mutation in the high-risk BC and OC genes BRCA1 or BRCA2. The combined effects of these variants on BC or OC risk for BRCA1 and BRCA2 mutation carriers have not yet been assessed while their clinical management could benefit from improved personalized risk estimates. Methods: We constructed polygenic risk scores (PRS) using BC and OC susceptibility SNPs identified through population-based GWAS: for BC (overall, estrogen receptor [ER]-positive, and ER-negative) and for OC. Using data from 15 252 female BRCA1 and 8211 BRCA2 carriers, the association of each PRS with BC or OC risk was evaluated using a weighted cohort approach, with time to diagnosis as the outcome and estimation of the hazard ratios (HRs) per standard deviation increase in the PRS. Results: The PRS for ER-negative BC displayed the strongest association with BC risk in BRCA1 carriers (HR = 1.27, 95% confidence interval [CI] = 1.23 to 1.31, P = 8.2 x 10(53)). In BRCA2 carriers, the strongest association with BC risk was seen for the overall BC PRS (HR = 1.22, 95% CI = 1.17 to 1.28, P = 7.2 x 10(-20)). The OC PRS was strongly associated with OC risk for both BRCA1 and BRCA2 carriers. These translate to differences in absolute risks (more than 10% in each case) between the top and bottom deciles of the PRS distribution; for example, the OC risk was 6% by age 80 years for BRCA2 carriers at the 10th percentile of the OC PRS compared with 19% risk for those at the 90th percentile of PRS. Conclusions: BC and OC PRS are predictive of cancer risk in BRCA1 and BRCA2 carriers. Incorporation of the PRS into risk prediction models has promise to better inform decisions on cancer risk management

Log odds of carrying an Ancestral Mutation in BRCA1 or BRCA2 for a Defined personal and family history in an Ashkenazi Jewish woman (LAMBDA)

INTRODUCTION: Ancestral mutations in BRCA1 and BRCA2 are common in people of Ashkenazi Jewish descent and are associated with a substantially increased risk of breast and ovarian cancer. Women considering mutation testing usually have several personal and family cancer characteristics, so predicting mutation status from one factor alone could be misleading. The aim of this study was to develop a simple algorithm to estimate the probability that an Ashkenazi Jewish woman carries an ancestral mutation, based on multiple predictive factors. METHODS: We studied Ashkenazi Jewish women with a personal or family history of breast or ovarian cancer and living in Melbourne or Sydney, Australia, or with a previous diagnosis of breast or ovarian cancer and living in the UK. DNA samples were tested for the germline mutations 185delAG and 5382insC in BRCA1, and 6174delT in BRCA2. Logistic regression was used to identify, and to estimate the predictive strength of, major determinants. RESULTS: A mutation was detected in 64 of 424 women. An algorithm was developed by combining our findings with those from similar analyses of a large study of unaffected Jewish women in Washington. Starting with a baseline score, a multiple of 0.5 (based on the logistic regression estimates) is added for each predictive feature. The sum is the estimated log odds ratio that a woman is a carrier, and is converted to a probability by using a table. There was good internal consistency. CONCLUSIONS: This simple algorithm might be useful in the clinical and genetic counselling setting. Comparison and validation in other settings should be sought