Molecular Pathways: Targeting RAC-p21-Activated Serine-Threonine Kinase Signaling in RAS-Driven Cancers

Cancers driven by oncogenic Ras proteins encompass some of the most deadly human cancer types, and there is a pressing need to develop therapies for these diseases. While recent studies suggest that mutant Ras proteins may yet be druggable, the most promising and advanced efforts involve inhibitors of Ras effector signaling. Most efforts to target Ras signaling have been aimed at the ERK mitogen-activated protein kinase and the phosphatidylinositol 3-kinase signaling networks. However, to date, no inhibitors of these Ras effector pathways have been effective against RAS mutant cancers. This ineffectiveness is due, in part, to the involvement of additional effectors in Ras-dependent cancer growth, such as the Rac small GTPase and the p21-activated serine-threonine kinases (PAK). PAK proteins are involved in many survival, cell motility, and proliferative pathways in the cell and may present a viable new target in Ras-driven cancers. In this review, we address the role and therapeutic potential of Rac and Group I PAK proteins in driving mutant Ras cancers

DNA methylation analysis of AKT1 promoter and HTR2A exon-I of Malaysian schizophrenia multiplex families with lower cognitive performance

Dysfunction of cognitive performance in schizophrenia has been associated with aberrant alteration of DNA methylation of several schizophrenia-risk genes. AKT1 and HTR2A are among the candidate genes for schizophrenia. Their expressions were found reduced in schizophrenia patients. Thus, we aimed to study the methylation status of AKT1 promoter and HTR2A exon-I in Malaysian schizophrenia patients and their affected family members. In this study, each participant was required to perform Trail Making Test (TMT) part A and B to measure their cognitive performance. Genomic DNA extracted from the peripheral blood of 12 Malaysian schizophrenia families and 12 controls families, was subjected to bisulfite conversion. The methylation status of CpG sites of AKT1 promoter at Chr14: 104796054 and HTR2A exon-I at Chr13: 46896918 were identified using methylation-specific polymerase chain reaction (MSP). Our results showed that schizophrenia patients performed worse in both TMT-A and B (p<0.0001) than healthy controls. The patients also displayed significantly (p=0.023) high level of methylation in AKT1 promoter compared to controls. Meanwhile, no significant difference (p=0.248) in methylation status was observed in HTR2A exon-I between schizophrenia and control groups. Therefore, methylation of AKT1 promoter in peripheral bloods of patients may involve in cognitive impairment and schizophrenia pathology. In addition, we were able to demonstrate the heritability of DNA methylation status across family members

Enhanced sensitivity of neutralizing antibody detection for different AAV serotypes using HeLa cells with overexpressed AAVR

A cell-based transduction inhibition assay (TI) is widely used in clinical trials to detect neutralizing antibody (NAb) titers against recombinant adeno-associated virus (rAAV), one of the most important criteria to exclude patients in gene therapy. Different cell lines are used in cell-based TI because the rAAV transduction efficiencies vary largely among serotypes. A cell line suitable for TI for most serotypes is highly desirable, especially for those with very low transduction efficiencies in vitro such as rAAV8 and rAAV9. Herein, we report an AAVR-HeLa, a stable cell line with overexpressed AAVR, a newly identified receptor for rAAVs, was established for cell-based TIs. The AAVR expression level in AAVR-HeLa cells was approximately 10-fold higher than in HeLa cells, and was stably transfected after twenty three passages. For all AAV serotypes (AAV1-10), except for AAV4, the transduction efficiencies increased significantly in AAVR-HeLa cells. It was demonstrated that the AAVR enhancement of transduction efficiency was only for rAAV and not for lentiviral and adenoviral vectors. According to the minimal multiplicity of infection (MOIs) for the assay, the NAb detection sensitivity increased at least 10 and 20 fold for AAV8 and AAV9, respectively. The seroprevalence of NAbs were investigated at the 1:30 level as a cutoff value using AAVR-HeLa cells. It was shown that the seropositive rate for AAV2 was 87% in serum samples from 99 adults, followed by lower seropositive rates for AAV5 (7%), AAV8 (7%) and AAV9 (1%). Venn diagram analysis showed the presence of cross-reactivity of NAbs to two or three serotypes in 13 samples (13.1%). However, no patient was found to possess NAbs for all the four serotypes. These results demonstrated that the AAVR-HeLa cell line may be utilized to detect the NAbs through cell-based TI assays for most of AAV serotypes

Towards a global partnership model in interprofessional education for cross-sector problem-solving

Objectives A partnership model in interprofessional education (IPE) is important in promoting a sense of global citizenship while preparing students for cross-sector problem-solving. However, the literature remains scant in providing useful guidance for the development of an IPE programme co-implemented by external partners. In this pioneering study, we describe the processes of forging global partnerships in co-implementing IPE and evaluate the programme in light of the preliminary data available. Methods This study is generally quantitative. We collected data from a total of 747 health and social care students from four higher education institutions. We utilized a descriptive narrative format and a quantitative design to present our experiences of running IPE with external partners and performed independent t-tests and analysis of variance to examine pretest and posttest mean differences in students’ data. Results We identified factors in establishing a cross-institutional IPE programme. These factors include complementarity of expertise, mutual benefits, internet connectivity, interactivity of design, and time difference. We found significant pretest–posttest differences in students’ readiness for interprofessional learning (teamwork and collaboration, positive professional identity, roles, and responsibilities). We also found a significant decrease in students’ social interaction anxiety after the IPE simulation. Conclusions The narrative of our experiences described in this manuscript could be considered by higher education institutions seeking to forge meaningful external partnerships in their effort to establish interprofessional global health education

A human PrM antibody that recognizes a novel cryptic epitope on dengue E glycoprotein

10.1371/journal.pone.0033451PLoS ONE74

p21-Activated Kinases Are Required for Transformation in a Cell-Based Model of Neurofibromatosis Type 2

NF2 is an autosomal dominant disease characterized by development of bilateral vestibular schwannomas and other benign tumors in central nervous system. Loss of the NF2 gene product, Merlin, leads to aberrant Schwann cell proliferation, motility, and survival, but the mechanisms by which this tumor suppressor functions remain unclear. One well-defined target of Merlin is the group I family of p21-activated kinases, which are allosterically inhibited by Merlin and which, when activated, stimulate cell cycle progression, motility, and increased survival. Here, we examine the effect of Pak inhibition on cells with diminished Merlin function.Using a specific peptide inhibitor of group I Paks, we show that loss of Pak activity restores normal cell movement in cells lacking Merlin function. In addition, xenografts of such cells form fewer and smaller tumors than do cells without Pak inhibition. However, in tumors, loss of Pak activity does not reduce Erk or Akt activity, two signaling proteins that are thought to mediate Pak function in growth factor pathways.These results suggest that Pak functions in novel signaling pathways in NF2, and may serve as a useful therapeutic target in this disease

Applications of proteomics: identification ofgenes associated with anti-cancer drug resistance, liver developmentand regeneration

abstractpublished_or_final_versionClinical OncologyDoctoralDoctor of Philosoph

A clinical guideline on the use of acustimulation on managing adult orthopaedic patients with postoperative nausea and vomiting

published_or_final_versionNursing StudiesMasterMaster of Nursin

Empowering Chinese abused women : a community-based participatory approach

Intimate partner violence (IPV) against women is a priority global health issue. Apart from the serious negative consequences for women’s health, IPV against women can lead to huge economic and health burdens on the community. In response to these serious health consequences and the high prevalence of IPV against women, interventions have been developed and demonstrated to have positive effects in preventing violence and addressing negative health outcomes. However, those interventions were theory-driven and developed in a top-down way based on an outside-expert approach, without women’s involvement and input. This approach is contrary to the principle of empowerment, which is the key of such interventions. A community-based participatory approach (CBPA) is adopted to empower Chinese abused women is this study. The objectives of this thesis are first to develop a culturally appropriate intervention for Chinese abused women using a CBPA based on such women’s expressed needs and the services and support they require. Through bringing women, professionals and community service providers together, a tripartite collaboration is established with engagement of the actual users throughout the process of development. The second objective is to assess the feasibility, acceptability and usefulness of an intervention developed under CBPA. A qualitative approach with three phases is adopted. Phase 1 was in the form of semi-structured focus group interviews with community-dwelling Chinese abused women in Hong Kong aimed at exploring their views of the services available, as well as their needs and preferences for services and support in response to violent victimisation. Phase 2 involved the use of CBPA to develop a needs-based, culturally appropriate empowerment intervention for women experiencing IPV. Phase 3 involved the implementation of the intervention and the use of semi-structured focus group and individual interviews to assess the feasibility, acceptability and usefulness of the Phase 2 intervention. In Phase 1, 11 abused Chinese women were interviewed in three focus groups. Conventional content analysis revealed that the women had a range of needs, personal, family and social, and their desires for services and support were multi-faceted and diverse. Personal needs were physical, emotional and concerned with parenting. As to family needs, the women were worried about family relationships; while they wanted informal social support on the social network side. The services and support the women wanted included mindfulness activity, relaxation and diversional therapy for stress reduction, whole-family activities (especially outdoors) to enhance family relationships, peer groups for sharing mutual support, and services to connect them to the community resources available. Three themes emerged from the focus group interviews in Phase 1, representing three levels of intervention: individual, family and community. In Phase 2, an integrated multi- component intervention was developed comprising a four-part, women-centred programme, “Women-centred, we are with you”, for Chinese abused women using CBPA. The intervention was based on the above-mentioned three levels, considered by the women to be the best model to address their complex and multi-faceted needs. In Phase 3, the “Women-centred, we are with you” programme was completed in four months, from October 2015 to January 2016. Afterwards, it was evaluated and the women’s experience of participating in CBPA explored; with 11 women interviewed in two focus groups and one as an individual. Conventional content analysis revealed that the women had high levels of satisfaction with the programme, which was well received. It was feasible to conduct the programme; the women showed high adherence, and women benefited from it in their well- being, family relationships and social networks. They gained a sense of achievement and their self-confidence was enhanced through participating in CBPA, which gave them more freedom and a sense of power and control over their own needs. The findings of this study suggest that abused women‘s needs should be assessed before any intervention so that an individualised and tailor-made type can be provided. Also, abused women had multi-faceted needs which should be addressed through multi-component interventions. The CBPA was also found to be a better approach for developing a tailor-made and culturally appropriate intervention for abused women. This is the first study to adopt CBPA in developing an intervention for community-dwelling Chinese abused women in Hong Kong, and there is evidence that participants found it acceptable, useful and feasible.published_or_final_versionNursing StudiesDoctoralDoctor of Nursin

Molecular Pathways: Targeting RAC–p21-Activated Serine–Threonine Kinase Signaling in RAS-Driven Cancers

Cancers driven by oncogenic Ras proteins encompass some of the most deadly human cancer types, and there is a pressing need to develop therapies for these diseases. While recent studies suggest that mutant Ras proteins may yet be druggable, the most promising and advanced efforts involve inhibitors of Ras effector signaling. Most efforts to target Ras signaling have been aimed at the ERK mitogen-activated protein kinase and the phosphatidylinositol 3-kinase signaling networks. However, to date, no inhibitors of these Ras effector pathways have been effective against RAS mutant cancers. This ineffectiveness is due, in part, to the involvement of additional effectors in Ras-dependent cancer growth, such as the Rac small GTPase and the p21-activated serine-threonine kinases (PAK). PAK proteins are involved in many survival, cell motility, and proliferative pathways in the cell and may present a viable new target in Ras-driven cancers. In this review, we address the role and therapeutic potential of Rac and Group I PAK proteins in driving mutant Ras cancers