Breast cancer is one of the most common diseases affecting women and approximately 1.3 million females are diagnosed each year with this disease worldwide. Breast cancer is a multi-factorial disease and it is difficult to predict or control the physiopathology. To date, one of the major risk factors, alongside the patient’s genetic background, is life time exposure to estrogen. Understanding the estrogen receptor (ER) has been a milestone in elucidating breast cancer biology, leading to advances in disease management. Alongside this, evidence from epidemiological studies suggests that dietary consumption of phytoestrogens may modulate disease progression. This study hypothesises that the interaction between some phytoestrogens (present in the pre-diagnosis diet or in the new diet adopted by breast cancer patients) and specific ER isoforms displayed in breast tumours influences the action of synthetic and endogenous estrogen in breast cancer cells. This study aimed to understand the interaction between estrogen, hormone drugs and phytoestrogens on the ER. In silico modelling of the ER focused on the wild type isoforms ERα and ERβ and different ligands (SWISS MODEL and docked through AutoDock Vina). Subsequently, isoforms of ERα and ERβ and different ligands (E1, E2, E3, PE, Tamoxifen, ICI 182,780) were modelled and tested by docking against the same set of ligands (E1, E2, E3, PE, Tamoxifen, ICI 182,780). The system described here highlighted the main amino acid residues of the LBD of ERα and ERβ along with ligand interactions for both agonists and antagonists, described in previous X-ray crystallography experiments. All of the phytoestrogens studied using AutoDock Vina interacted with the hormone binding site of both ERα and ERβ, due the phenolic ring of the studied structure which favoured the interaction with the hydrophobic environment of LBD amino acids. All of the dietary phytoestrogens showed lower binding affinity (< 9.1 Kcal/mol) compared with estradiol (-10.6 Kcal/mol) in all the isoforms and isotypes studied, suggesting that phytoestrogens should not displace estradiol from the LBD, however it remains unclear if PE can act as an agonist compound in the ER pathways. Also, some phytoestrogens appeared to have greater affinity to the ERα and ERβ than Tamoxifen (antagonist models), but it is uncertain as to whether the resulting structure will interfere with subsequent interactions. Further laboratory experiments will be necessary to understand the impact of the PE in the ERs structure and the respective role in the ER pathway. The data from this computer modelling approach has provided an insight into the interactions between endogenous estrogens, drugs, phytoestrogens and ER. The in silico studies generated a system that recapitulated data obtained by other research groups (experimentally) and will be of value as a screening tool for further studies of new drugs and exogenous estrogens and their potential role in ER-induced breast cancer pathophysiology.

Background: Coronary Heart Disease (CHD) is one of the leading causes of death in both men and women worldwide. It is well documented that early diagnosis and treatment of CHD is associated with better outcomes. This has led to the establishment of targets to ensure prompt access to services for potential CHD (e.g. Rapid Access Chest Pain Clinics). Research has shown that these public health targets have caused decreases in morbidity and mortality rates for CHD in the UK. However, despite these improvements health services are still limited by help-seeking practices of patients as they can only act once a patient has presented for treatment. A number of studies have explored the reasons why patients delay help-seeking for CHD symptoms in an emergency context (i.e. having a heart attack). Many studies have focused on gender and have often suggested that women with emergency CHD symptoms delay help-seeking, although this is controversial. Other studies have suggested help-seeking delay is influenced by multiple intersecting factors (e.g. age, ethnicity and contextual influences) and not just gender. No studies have examined help-seeking for suspected CHD symptoms in the context of accessing Rapid Access Chest Pain Clinics (RACPC). Given the lack of understanding in this area, an explorative qualitative study was undertaken to answer the research question: what are the help-seeking experiences of men and women referred to a rapid access chest pain clinic? Methods: A total of 30 men and women with a range of ages and ethnicities referred to a RACPC for the investigation of their symptoms were enrolled in this study. Participants took part in semi-structured interviews that focused on attribution of symptoms and how that, amongst other things, influenced help-seeking decisions for their symptoms. The data was analysed thematically to explore men’s and women’s experiences and the help-seeking decisions they made. The study findings and relevant literature were used to inform the development of a patient information leaflet to assist with recognition of potential CHD symptoms and to promote help-seeking. Results: The study found, in general, that: attribution of symptoms was linked to contextual factors; reluctance to seek help and response to symptoms contributed to delay; the influence of others acted as enablers of help-seeking; and barriers were linked to accessibility of GP services and time off work. Additionally, some participants had mixed reactions to a negative diagnosis at the end of RACPC assessment (i.e. symptoms not of CHD origin). Some participants expressed frustration at not having an answer for their symptoms, whereas others said they felt like a ‘fraud’ for wasting NHS resources. Not all participants had negative reactions and many were delighted that their symptoms were not heart-related. When it came to perceptions of risk of CHD, most believed the ‘male lifestyle’ was more risky and therefore increased CHD risks in men, but that increasingly women were living ‘male-like lifestyles’ (e.g. working full time, smoking, drinking and eating a poor diet), thus increasing their risk of CHD. Conclusion: This novel study based in the RACPC context has produced important findings in this previously unexplored area. Earlier qualitative research based in the emergency CHD context has highlighted the challenges around symptom attribution, attitudes to help-seeking and response to symptoms, and how these factors contribute to delay. This current study showed that there were many similarities between the two different contexts (emergency and non-emergency). These findings can be used to produce health promotion literature to encourage early help-seeking for non-emergency CHD in the RACPC context in both men and women. The output of the current research makes a contribution to practice in my profession through the development of a lay patient resource to promote help-seeking in the RACPC context.

Thrombotic thrombocytopenic purpura (TTP) is a rare, life-threatening disease with a reported incidence of 6 cases per million per year in the UK. It is characterised by episodes of microangiopathic haemolytic anaemia and thrombocytopenia, with the widespread presence of platelet-rich thrombi in the microcirculation, leading to end-organ damage. TTP is a clinically heterogeneous disorder caused by autoantibody inhibition or clearance or by a deficiency in activity or secretion of the von Willebrand factor cleaving protease (ADAMTS13). Over 100 mutations have been identified in ADAMTS13 yet, in some cases, these mutations in congenital TTP alone do not explain the disease phenotype, particularly in late-onset congenital TTP. One aim of this study was to analyse the effect of single nucleotide polymorphisms (SNPs), in conjunction with ADAMTS13 mutations, which have been identified in a cohort of TTP patients with varied clinical phenotype. Twenty mutant expression plasmids containing an ADAMTS13 mutation and/or SNP have been constructed by site-directed mutagenesis (SDM). Fourteen plasmids contained a new non-synonymous mutation (C3484T) which has been rectified by further SDM. After full sequencing of the plasmid ADAMTS13 insert, 24 clones were identified as viable and the resultant protein was expressed in HEK293T cells and analysed by Western blot and ADAMTS13 antigen ELISA. Blots were scanned by densitometry and the intensity of the protein species corresponding to ADAMTS13 determined using ImageJ. The predicted effects of mutations and/or SNPs were annotated by the in silico computational tools SIFT, PolyPhen2, I-Mutant 2.0 and SNPEffect 4.0.Results from this genotype study were compared to clinical phenotype by correlating data held within the TTP Registry at University College London. The resultant analysis showed the mutations located in the N-terminal end of ADAMTS13 protein induced more secretory defects and SNPs had less of an effect on phenotype. The p.R7W and p.A1033T SNPs showed more of an effect on secretion of ADAMTS13 when the mutation was located in the Cterminal end (p.R1060W) which is associated with late-onset phenotype. However, other environmental factors such as changes in the ADAMTS13-VWF axis during pregnancy and infection make a correlation between genotype and phenotype in TTP more challenging.

Emerging zoonotic viruses pose an increasing threat, causing outbreaks with high rates of morbidity and mortality and frequently significant economic implications. Often, there is a lack or shortfall of effective prophylaxis and diagnostic capabilities. Research towards their development, together with improved surveillance activities are high priority activities to prepare and respond to outbreak threats. Yet handling these viruses commonly requires high containment levels. This can be circumvented by the use of replication defective pseudotyped viruses (PVs), incorporating the viral envelope protein of interest which constitutes the primary surface antigen. This permits the serological detection of neutralising antibodies without the need to handle live virus, as well as other viral entry studies. Hence, PVs are increasingly proving to be a valuable tool for emerging virus research. The aim of this study was to exploit novelties in the unique flexibility of the PV platform to allow the serological assessment of emerging viruses and evaluate technical aspects towards standardisation. Current prophylaxis provides robust protection against rabies virus, yet only confers limited protection against other lyssavirus species, which have a near 100% fatality rate. It is thought protection is afforded against isolates of phylogroup I rabies virus, yet there is limited biological data for the Arctic-like rabies virus (AL RABV) lineage which is endemic across the Middle East and Asia. Although other lyssaviruses pseudotype efficiently, titres of AL RABV PV were low. Within this study, high titre PV was produced by constructing chimeric envelope proteins, splicing the AL RABV ecto-transmembrane domain with the cytoplasmic domain of vesicular stomatitis virus. Comparisons showed this did not alter the serological profile of the AL RABV and they were effectively neutralised by vaccines and antivirals. It could therefore be concluded that they do not pose a significant public health risk. However it is recognised broadly neutralising prophylaxis needs to be developed to protect against more divergent lyssaviruses. In a further study, again utilising the flexibility to manipulate the envelope protein, PV was produced switching the five known antigenic sites of the envelope protein between a phylogroup I (rabies virus) and III (West Caucasian bat virus) isolate. Screening polyclonal sera via a neutralisation assay, the immunologically dominant sites for phylogroup I and III were identified as III and I respectively. This can act to inform future development of more broadly neutralising vaccines. The 2013-16 outbreak of Ebola virus focused global efforts towards the urgent need for effective vaccines and antivirals. To permit low containment level serology studies to assist their development, a panel of filovirus PVs were rapidly produced. Work was carried out to optimise their method of production; determining lentiviral core PV produced by transfecting HEK 293T/17 cells was most efficient. Efforts to repeat the use of chimeric envelope proteins to increase titre proved unsuccessful. The evaluation of target cell lines permissive to infection and appropriate for neutralisation assays identified that the CHO-K1 cell line produced the clearest data. The PV neutralisation assay was subsequently applied to a range of projects to assess candidate prophylaxis and demonstrated the value of the platform to respond to emerging virus outbreaks. Given the increasing prominence in the use of PV, work was undertaken to expand their utility and methods for standardisation. An assessment of new reporter genes found a red fluorescent protein, with a nuclear localisation signal, improved the clarity of data collection and output in additional spectrum to the current repertoire. To be able to correlate the disparate readout units of fluorescent and luminescent reporters, recorded as infectious units (IFU) and relative light units (RLU) respectively, a new construct was produced to integrate and equally express two reporters from cells transduced with PV. It was determined that approximately 1260 RLU equates to 1 IFU, although future work to determine how this fluctuates between cell lines is required. Finally, alternative methods to quantify PV were evaluated, measuring the number of particles, genome copies and reverse transcriptase (RT) activity, in addition to the currently used biological titre. It was found that measures of genome copies and RT activity, in combination with biological titre provides information on the quality of PV preparations and could be used to standardise assay input.

Introduction Nutrition can impact significantly on cancer occurrence, recurrence and survival. Many people affected by cancer seek individualised nutrition advice, but globally this remains an unmet need for many. Nutritional therapy (NT) practitioners provide personalised nutritional care, including complementary care for people affected by cancer. Until now, there is little documentation internationally of NT practice with people affected by cancer. This mixed methods study explores NT practitioners’ perspectives, to inform professional development needs to ensure safe evidence-based practice with people affected by cancer. Methods An on-line anonymised survey was open to all UK registered NT practitioners. Data was collected on practitioner characteristics, patterns of practice with people affected by cancer and perceived barriers to that practice, the use of research evidence in practice, and practitioners’ perceived needs for training and support. Data on practitioner characteristics were compared between practitioners who work with people affected by cancer (CP) and those who do (NCP). Preliminary themes emerging from the survey informed the second phase, in which semi-structured individual interviews (n=21) and one group interview were used for in depth exploration of practitioner perspectives on practice with people affected by cancer. Topics explored included rewards and motivation, barriers and challenges to practice, and professional development needs. Interview transcripts were analysed thematically, using Framework methodology. Results 274/888 (31%) UK registrants completed the survey. CPs were more likely than NCPs to have additional health and life science (HLS) qualifications (p=0.009) and to have been in practice for longer (practice for more than 10 years, p=0.012). 80% survey respondents overall, including CPs, worked alone for at least some of the time. Practitioners experienced reward in empowering people affected by cancer to make dietary changes, for some their own cancer journey or witnessing cancer in a loved one was a motivator. Significant challenges and barriers to practice included managing the emotional and psychological demands of working with people affected by cancer, lack of recognised specialist training and resources, and lack of engagement with or perceived opposition from oncologists. To overcome these barriers, professional development needs identified were firstly specialist clinical supervision, mentoring and networking to overcome isolation and share best practice. Secondly, specialist training and resources to facilitate accessing, critically appraising and communicating evidence in practice are required. Thirdly, engagement with mainstream professionals working in cancer care requires improvement. Conclusions This is first detailed exploration of NT practitioner perspectives on working with people affected by cancer and the data have revealed important areas for developing training and support. Findings are informing development of frameworks for practitioner support including clinical supervision and specialist training, to support safe evidence-based practice and better communication and integration with mainstream cancer care. Future exploration of NT practitioners’ perspectives should assess the impact of proposed professional developments highlighted here. Further exploration of perspectives of mainstream healthcare professionals with whom nutritional therapy practitioners may engage is also recommended.

This qualitative study explores the personal and professional journey of a group of healthcare professionals who undertook higher education (HE) or training in social innovation and put that learning into practice. Social Innovation is a relatively new concept in the field of health, and research is, in the main, confined to the structural and organisational aspects of innovation. Studies looking at the personal development of the innovators are rare, and no study has researched first-hand the experiences of new healthcare innovators as they learn how to set up and manage their own projects. Twenty-six community health professionals were interviewed. The participants were selected because they had either attended a HE programme or independent training on social innovation and were interested in setting up their own innovation in their clinical practice setting. Individual interviews were achieved using Skype which proved to be an effective data collection method and allowed for a geographically dispersed sample. Thematic Analysis allowed several key themes to emerge from the data: the importance of personal resilience; increase in confidence; how levels of self- efficacy played a key role in their success; learning to shift from working in glorious isolation to seeking help from influential others. Improved technical skills and becoming better organised were also powerful factors. However, one finding proved pivotal to their success - finding themselves. The majority talked of discovering the ‘real me’ as a result of their learning, mixing with likeminded others and the first-hand experience of the struggle of developing a project, often in the face of opposition. For many, the positive changes transferred to their personal lives. The findings suggest that many community health professionals have an inner drive to improve their clinical practice, but do not always know the best way to do this without formalised help. The educational input enhanced their learning and also impacted on their personal development enabling them to proceed with their innovations. These findings are supported by research in the broader field of industry indicating that, whatever the context, there is a commonality of spirit, an ability to persevere and overcome adversity among innovators. These findings are therefore generalisable to others contemplating innovative projects in health and social care settings. In addition, the health and social professional curricula will benefit from including the subject of innovation within their educational programmes and subsequently staff and managers who work with innovative practitioners will also benefit from working with innovative professionals.

Non-alcoholic fatty liver disease (NAFLD) is a metabolic disease characterised by the accumulation of fat in the liver. It is estimated that 33 % of the UK population have NAFLD with 2-5 % progressing to non-alcoholic steatohepatitis (NASH). Due to a lack of an outright therapy for NAFLD, treatment has been mainly focussed on managing the conditions associated with the disease such as obesity, diabetes mellitus and hyperlipidaemia. This study aimed to investigate the means by which hepatocyte protection is conferred by Gentiana plants (Gentiana lutea, Gentiana macrophylla, Gentiana scabra and Gentiana rigescens) used in herbal medicine for the management of non-alcoholic fatty liver diseases (NAFLD). The role played by some of the inherent Gentiana phytochemicals including: gentiopicroside, sweroside and swertiamarin in promoting hepatocyte protection against the cytotoxic effects of fatty acids were also investigated. Gentiana species: lutea, macrophylla, rigescens, and scabra are known to protect and enhance hepatocyte viability via their antioxidant, anti-inflammatory and bitter components including: amarogentin gentianine, iso-orientin, swertiamarin, gentiopicroside, and sweroside. This study was necessitated due to a lack of adequate research on the hepatoprotective effects of the above-named Gentiana species and phytochemicals with special emphasis on their effect on mitochondrial respiration in the presence of fatty acids. At the time of submission, this was the first study to utilise the seahorse mitochondria stress assay to investigate the Gentiana species as well as phytochemicals: gentiopicroside, sweroside and swertiamarin. It was also found that the most abundant phytochemical in all four Gentiana species was gentiopicroside (up to 4.6% g/g), followed by swertiamarin (0.21–0.45% g/g), and sweroside (0.03- 0.4 % g/g). Furthermore, it was also observed that the methanolic extracts of all four Gentiana protected HepG2 and THLE-2 cells by inhibiting arachidonic acid from diminishing cell replication but showed a mitogenic effect mostly observed in gentiopicroside, Gentiana lutea and Gentiana macrophylla. It was concluded that phytochemicals: gentiopicroside, sweroside and swertiamarin play key roles in the hepatocyte protection exerted by methanolic extracts of Gentiana lutea, Gentiana macrophylla, Gentiana scabra and Gentiana rigescens against the cytotoxic effects of fatty acids. This protection is conferred by enhancing mitochondrial function in terms of increasing maximal respiratory capacity in response to high influx of fatty acids, promoting ATP production as well as scavenging ROS produced as a result of high fatty acid influx and increased mitochondrial respiration. However, the mitogenic effect observed in gentiopicroside and Gentiana macrophylla requires further studies using unmodified primary hepatocytes to gain better understanding.

Background: Alcohol and Free fatty acids such as palmitate are known to promote liver injury. However less mechanistic information is available regarding omega fatty acids ratios with/out alcohol. In healthy populations omega 6/3 ratios are between 1:1 to 4:1, whereas high ratios ( > 15:1) are thought to correlate with the pathogenesis of fatty liver disease. This study aimed to investigate liver lipotoxicity and mitochondrial dysfunction due to imbalanced omega 6/3 ratios alone or in the presence of alcohol. Method: Human hepatoma cell line, VL-17A cells were treated with individual fatty acids (Palmitic (PA), Stearic (SA), Linoleic (LA), alpha-Linolenic (ALA), Arachidonic (AA) and Docosahexaenoic (DHA) acids) with various concentrations ranging between 0.5 μM to 300 μM and omega 6/3 ratios (1:1, 4:1, 15:1 and 25:1) with/out alcohol (100 mM) for 24, 48 and 72 h after which lipid accumulation and cell toxicity was assessed. Subsequent studies at 24 hr examined oxidative stress, mitochondrial function and lipogenic proteins. Results: PA treatment showed a detrimental effect on cell viability and lipid accumulation than SA after 48 h and 72 h (P < 0.05); omega 3 (ALA and DHA) did not show any significant effect; LA exhibited a significant reduction in cell viability at 24 h only without causing any change in lipid accumulation; AA treatment significantly reduced cell viability at 48 h and 72 h (P < 0.05) but only showed a significant lipid accumulation after 48 h. AA/DHA (omega 6/3) ratios (15:1 & 25:1) caused a significant reduction in cell viability after 24 h, 48 h and 72 h (P < 0.001) and only (15:1) ratio inversely corresponded to an elevation in lipid level after 24 h and 48 h (P < 0.01). At 24 hr treatment, high AA/DHA ratio of 25:1 led to increased expression of stearoyl-CoA desaturase (SCD1) and decreased peroxisome proliferator activated receptor alpha (PPARα) expression at 15:1 and 25:1 (P < 0.05), while all ratios showed a significant decrease in the cannabinoid receptor (CB2) expression compared to control but actually increased when compared to the 1:1 ratio. Although the expression of CB1 was slightly increased (P > 0.05), sterol regulatory element-binding protein 1 (SREBP1) did not show any change. AA/DHA ratios also showed a significant decrease in ATP production (P < 0.01), basal respiration, maximal respiration and spare mitochondrial capacity and this effect was greater with high ratios (P < 0.001). Reactive oxygen species (ROS) production increased significantly, particularly with high AA/DHA ratios (15:1 and 25:1) (P < 0.001) alone and in the presence of alcohol (P < 0.01). Conclusion: The data suggests that lipid accumulation and toxicity occur with saturated and unsaturated fatty acids and high omega 6/3 ratios. The latter possibly due to the pro-inflammatory products of AA. This study confirms that high AA/DHA ratios with/out alcohol increase ROS production and high AA/DHA alone induce mitochondrial dysfunction and increase lipogenesis pathways by activating lipogenic factors causing steatosis and consequently promoting the development of fatty liver disease. Further work aims to elucidate the effect of fatty acid/alcohol on lipid synthetic and endocannabinoid pathways, which will further our understanding of fatty liver disease development.

The laboratory diagnosis of Strongyloides stercoralis (S. stercoralis) at the Department of Clinical Parasitology (DCP) by the routine methods of microscopy and Strongyloides culture is not sensitive due to the, usually, low parasite load and intermittent larval excretion of the parasite. Serology (enzyme-linked immunosorbent assay) suffers from a lack of specificity because Strongyloides antibodies are known to cross- react with schistosomal, filarial and other helminthic antibodies in serological tests. Moreover, antibody levels are slow to decline after successful treatment therefore serology cannot be used to monitor point of cure. A missed diagnosis of strongyloidiasis in immunocompromised patients or those about to undergo iatrogenic immune suppression may have severe, even fatal, consequences. The disease is poorly studied because of the lack of sensitive, specific and cost-effective tests. Therefore, the decision was made to evaluate and validate nucleic acid amplification techniques (NAATs) for the diagnosis of S. stercoralis for use in a well- resourced specialist referral parasitology laboratory. A novel loop mediated isothermal amplification (LAMP) assay was also developed for use in resource- limited regions. The study was conducted over two years (2014-2016) and examined 284 residual diagnostic samples. The cohort was drawn from patients attending a central London western travel medicine (WTM) clinic. The NAATs chosen for this study were a published real- time PCR (qPCR) assay (ten Hove et al., 2009) and a novel LAMP assay. The NAATs were compared to the combined reference standard of microscopy, culture and serology for the diagnosis of S. stercoralis in stool samples. The development of the novel LAMP assay for use in resource- limited areas included the investigation of methods for rapid, simple and cost- effective DNA extraction. The qPCR and LAMP assays detect target DNA within areas on either side of the S. stercoralis 18S rRNA genome hypervariable region (Hasegawa et al., 2009). In this study the LAMP and qPCR assays demonstrated a limit of detection of 10-3 and 10-4, respectively for S. stercoralis DNA detection in clinical samples. Specificity was determined for the LAMP and qPCR assays to be 100% and 94.83%, respectively and the cost per test was calculated as £4.80 and £8.21, respectively. In this study, persistence of S. stercoralis DNA in clinical samples was improved when the samples were stored at -20oC. While the LAMP assay has a shorter turnaround time and is less costly than qPCR, the superior efficiency of qPCR detection of S. stercoralis DNA in clinical samples established that the qPCR assay was a more suitable addition to the diagnostic repertoire at a high- throughput WTM clinic. The LAMP assay showed promise for deployment in resource- limited areas and as a point- of- care test but further work is required to optimise the LAMP assay for these purposes.

Signalling through the Frizzled-dependent planar polarity pathway is a conserved mechanism that polarises cells in a plane perpendicular to the apical-basal axis. Epithelial cells in many organisms exhibit planar polarity, including the Drosophila wing and eye, and during convergent extension movements in vertebrates. The core planar polarity proteins involved in establishing planar polarity exhibit asymmetric subcellular localisations. Currently little is known about how the asymmetry is generated and maintained. This work uses techniques such as live imaging and FRAP of Drosophila epithelial tissues to study protein turnover, to understand these processes. I focused on the core proteins Frizzled (Fz) and Flamingo (Fmi). Both proteins are localised at apical junctions in membrane subdomains (puncta). FRAP on puncta of either protein shows there is a large immobile fraction compared to surrounding junctional regions. Mutations in the other core proteins result in a loss of Fz and Fmi protein asymmetry, a loss of large puncta, as well as a reduction in the size of the immobile fraction. My data support a model where the six known core proteins are mutually stabilised by each other into large puncta where they form intercellular asymmetric junctional complexes. These puncta represent groups of aligned intercellular protein complexes, which I propose are required for establishing and coupling cellular asymmetry. The roles of planar polarity during coordinated cell rearrangements have not been extensively studied in Drosophila. This work shows that the core proteins are required for coordinated cell rearrangements during Drosophila embryonic tracheal cell intercalation. Regulation of the adhesion protein DE-cadherin is required for cell intercalation and this is under control of the core proteins via a RhoGEF2-dependent mechanism. Therefore this work supports a role for the core proteins in regulating cell adhesion during cell movements.