Correlates of substance use in a large naturalistic cohort of young people with early and emerging psychosis.

BACKGROUND: Substance use remains a barrier to recovery for young people accessing early intervention services for psychosis. While correlates of use have been explored in populations experiencing a first episode of psychosis (FEP), sample sizes have been small and less research assesses cohorts at ultrahigh risk of psychosis (UHR). METHODS: This study uses data from a naturalistic cohort including UHR and FEP participants (N = 1252) to elucidate clinical correlates of use in the past 3 months of any illicit substance, amphetamine-type stimulants (ATS), cannabis, and tobacco. Moreover, network analysis based on use of these substances and additionally alcohol, cocaine, hallucinogens, sedatives, inhalants, and opioids was completed. RESULTS: Young people with FEP used substances at significantly higher rates than those at UHR. High concurrence of use was seen between substances. In the FEP group, participants who had used any illicit substance, ATS, and/or tobacco had increased positive symptoms and decreased negative symptoms. Young people with FEP who used cannabis had increased positive symptoms. In the UHR group, participants who had used any illicit substance, ATS, and/or cannabis in the past 3 months showed decreased negative symptoms compared to those who had not. CONCLUSION: A distinct clinical picture of more florid positive symptoms and alleviated negative symptoms seen in those who use substances in the FEP group appears muted in the UHR cohort. Treating young people at UHR in early intervention services represents the earliest opportunity to address substance use early to improve outcomes.

Social inclusion, intersectionality, and profiles of vulnerable groups of young people seeking mental health support.

BACKGROUND: headspace centres provide enhanced primary mental healthcare for young people. A priority is to provide services for all young people irrespective of a range of social disadvantages or social exclusion. The aims of this study were to: (i) delineate extent of social inclusion across domains of housing, studying/employment, functioning, alcohol, and other drug use; and (ii) map profiles of young people deemed vulnerable to experiencing additional barriers to accessing services based on their social inclusion domains (e.g., those living in unstable housing, not in employment/education, and/or experiencing intersecting or multiple forms of disadvantage or difficulties), including detailing their clinical characteristics. METHODS: Young people were recruited from five headspace centres. Data relevant to social inclusion were examined. Multivariate logistic regression models were used to determine overlap between vulnerable groups, functional, social, clinical, and behavioural factors. RESULTS: 1107 young people participated, aged 12-25 years (M = 18.1 years, SD = 3.3), most living in stable housing (96.5%) and engaged in studying/employment (84.8%). Specific vulnerabilities were evident in young people with NEET status (15.2%); in unstable accommodation (3.5%); of culturally diverse backgrounds (CALD) (12.2%); living in regional areas (36.1%); and identifying as lesbian, gay, bisexual, transgender, intersex, queer/questioning, and asexual plus (LGBTIQA+; 28.2%). Higher levels of distress, substance use, functional impairment, and lower social support were reported by those who were NEET and/or in unstable housing. LGBTIQA+ status was associated with high distress, depressive symptoms, and suicidal ideation. CONCLUSIONS: Most participants reported good social support, stable housing, and engagement in work or education. Those deemed vulnerable were likely to experience social exclusion across multiple domains and reported more mental health problems. The co-occurrence of mental ill-health and social exclusion highlights the importance of integrated mental healthcare.

The psychometric characteristics of the Kessler Psychological Distress Scale (K6) in help-seeking youth: What do you miss when using it as an outcome measure?

This is the first study to describe psychometric properties of the Kessler Psychological Distress Scale (K6) in a large cohort of help-seeking young people presenting to primary mental health care services. The aim was to determine whether the K6 was appropriate for monitoring outcomes in such settings. 1067 young people were recruited from Australian headspace services. We examined dimensionality of the K6, measurement invariance, and how the K6 correlated with the the Patient Health Questionnaire-9 (PHQ-9)and the Generalised Anxiety Disorder-7 Scale (GAD-7). Standardised Response Mean (SRM) and Cohen's d effect size (ES) were used to examine 3-month stability of the K6. The best-fitting model was a two-factor model: (i) nervous and restlessness; and (ii) hopeless, worthless, depressed and effort. Measurement non-invariance was observed for sex and age groups. K6 strongly correlated with the PHQ-9 and GAD-7. The K6 was less sensitive to change compared to these other two measures. There was some support for the K6 being a screener for young people presenting to primary care; however, there issues arise with its use as an outcome measure. These issues include measurement non-invariance, concern about the dimensionality and focus of items, and its sensitivity to change.

Clinical and functional characteristics of a subsample of young people presenting for primary mental healthcare at headspace services across Australia.

PURPOSE: Headspace services provide treatment options to young people seeking mental healthcare. To obtain a better understanding of needs and characteristics of this population, and effectively evaluate services, we require novel youth-specific outcome measures. As part of our broad research program to establish such measures, a sample of young people were recruited and assessed. The study describes (i) methodology used to obtain clinical, functioning, and substance use characteristics of young people presenting to headspace services; and (ii) an overview of these characteristics. METHODS: Young people presenting to headspace centres were recruited. Multidimensional information was obtained relating to clinical and functional outcomes, demographic information, and lifestyle factors. RESULTS: 1107 young help-seeking individuals were recruited. Participants were most likely young adults aged M = 18.1 years, SD = 3.3, with diagnoses of depression and/or anxiety (76.6%, n = 801), engaged in work and study (84.9%, n = 890), and living with parent(s) (68.9%, n = 736). Impairments in functioning were moderate as indicated by the Social and Occupational Functioning Assessment Scale (M = 65.2, SD = 9.5), substance use was common (alcohol 62.7%, n = 665; illicit substances 30.5%, n = 324), and current suicidal ideation was reported by a third (33.6%, n = 358). CONCLUSIONS: A broad dataset was obtained providing an insight into key clinical, functional and quality of life characteristics of these individuals. We observed that young people present with complex problems, comorbid diagnoses, moderate levels of symptomatology, impairments in functioning, substance use, and suicidal ideation. This work provides the foundation for our broader research program aiming to develop novel, relevant and youth-specific, change and outcome measures.

Awareness of headspace youth mental health service centres across Australian communities between 2008 and 2015.

Background: Two common barriers to help-seeking are lack of awareness of appropriate services, and low mental health literacy. The headspace awareness campaigns are designed to address these factors.Aims: To examine whether distance from a headspace centre affects community awareness of headspace, and whether general awareness of headspace changed between 2008 and 2015.Method: Responses from 4707 participants aged 12-25 years, collected in 2008 and 2015, were analysed. The effect of headspace centre location on awareness of services was assessed by comparing awareness between those living in headspace areas (within 20 km of a centre) and those who were not. Change in awareness between 2008 and 2015 was assessed.Results: Awareness of headspace and its services was significantly greater among those living in headspace areas than among those living further away. Within headspace areas, awareness increased by 27% between 2008 and 2015. Prompted and unprompted awareness were significantly greater in 2015 than in 2008.Conclusions: Awareness of headspace has increased over time; however, innovative awareness campaigns are needed for those residing in non-headspace areas. Continued funding to increase headspace's national coverage, improving mental health literacy and service access for youth and their families, particularly those living in non-headspace areas, is needed.

Including a client sexual health pathway in a national youth mental health early intervention service--project rationale and implementation strategy.

Young people have higher rates of sexually transmissible infections (STIs) than the general population. Research has shown that there is a clear link between emotional distress, depression, substance abuse and sexual risk taking behaviours in young people. 'headspace' is a youth mental health early intervention service operating in more than 55 locations around Australia. This setting is ideal for accessing troubled young people who are at risk of STIs and are unlikely to attend mainstream general practice services. In 2013, a partnership was formed between headspace National Office and Hunter New England Local Health District to develop a clear clinical pathway to ensure at-risk headspace clients received sexual health assessments, advice, appropriate treatment and referral as indicated. This article describes the implementation of the pathway at the national level and a control trial to measure its efficacy. Future work will involve providing sustainable sexual health training for headspace general practitioner and allied health professionals to supplement the roll-out of the pathway in headspace centres across Australia. Support will also be provided in adapting headspace environments to encourage service utilization by Aboriginal and Torres Strait Islander and gay, lesbian, bisexual, transsexual and intersex young people.

Functional characterization of cancer-associated Gab1 mutations.

Grb2-associated binder 1 (Gab1) is a docking protein that transduces signals from a variety of tyrosine kinases, including Met and the epidermal growth factor receptor (EGFR). Although the related protein Gab2 is strongly implicated in human cancer, a role for Gab1 has been less clear. However, a screen for gene mutations in breast cancer identified two somatic mutations in Gab1, Y83C and T387N. In this paper we describe the functional characterization of these Gab1 mutants. MCF-10A immortalized mammary epithelial cells overexpressing Gab1 Y83C and T387N exhibited a more elongated, fibroblastic phenotype compared with wild-type Gab1 controls. Expression of Gab1 or the mutants promoted epidermal growth factor (EGF)-independent proliferation in monolayer culture to a similar degree. However, in Matrigel culture, both mutants enhanced the formation of acini exhibiting an aberrant, branched morphology. In addition, expression of the mutants modestly increased Erk activation. The two mutants also enhanced branching morphogenesis in a different mammary epithelial cell line, HC11. To gain further insights into the mechanism of action of these mutations, we mapped Gab1 phosphorylation sites by mass spectrometry. This detected phosphorylation of T387 but ;not Y83. Cellular stimulation with EGF or hepatocyte growth factor (HGF) led to a transient, or sustained, induction of T387 phosphorylation, respectively. As T387 corresponds in position to Gab2 T391, which suppresses Gab2 signaling in a phosphorylation-dependent manner, these data support a model in which the T387N mutation abrogates negative-feedback regulation of Gab1. Interrogation of publically-available databases revealed additional cancer-associated mutations at, or in close proximity to, identified serine/threonine phosphorylation sites in other docking proteins. These data indicate that aberrant Gab1 signaling can directly contribute to breast cancer progression, and that negative feedback sites in docking proteins can be targeted by oncogenic mutations.

Preclinical evaluation of dual PI3K-mTOR inhibitors and histone deacetylase inhibitors in head and neck squamous cell carcinoma.

BACKGROUND: We examine the potential value of a series of clinically relevant PI3K-mTOR inhibitors alone, or in combination with histone deacetylase inhibitors, in a model of head and neck squamous cell carcinoma (HNSCC). METHODS: Head and neck squamous cell carcinoma cell lines, human keratinocyte and HNSCC xenograft models were treated with histone deacetylase inhibitors (HDACIs) and new generation PI3K and dual PI3K-mTOR inhibitors either alone or in combination. Cell and tumour tissue viability and proliferation were then determined in vitro and in vivo. RESULTS: Phosphatidylinositol-3-phosphate kinase, AKT and dual PI3K-mTOR inhibitors caused marked in vitro enhancement of cytotoxicity induced by HDACIs in HNSCC cancer cells. This effect correlates with AKT inhibition and is attenuated by expression of constitutively active AKT. Histone deacetylase inhibitor and phosphatidylinositol-3-phosphate kinase inhibitors (PI3KIs) inhibited tumour growth in xenograft models of HNSCC. Importantly, we observed intratumoural HDAC inhibition and PI3K inhibition as assessed by histone H3 acetylation status and phospho-AKT staining, respectively. However, we saw no evidence of improved efficacy with an HDACI/PI3KI combination. INTERPRETATION: That PI3K and dual PI3K-mTOR inhibitors possess antitumour effect against HNSCC in vivo.

Indole-3-carbinol - induced growth inhibition can be converted to a cytotoxic response in the presence of TPA+Ca(2+) in squamous cell carcinoma cell lines.

We examined the possibility that I3C, when combined with a differentiation stimulus (TPA+CaCl(2)), would sensitise SCC cells to a differentiation stimulus. We report that I3C induces a profound growth inhibition in SCC cells that is dissimilar to the growth inhibition required to initiate differentiation. Moreover, we report that I3C, when combined with TPA+CaCl(2) treatment, induces a loss of colony forming ability that was differentiation and senescence - independent but was due to delayed cytotoxicity. This study shows that I3C in combination with a PKC activator+Ca(2+) may be a useful therapeutic strategy for treating oral SCC.

Transfection of human peripheral blood mononuclear cells using immunoporation.

Immunoporation has been found to be able to efficiently transfect a wide range of human cultured cell lines. This report shows that peripheral blood mononuclear cells can also be efficiently transfected using immunoporation. The immunoporation of the cells with fluorescent TMR-Dextran, using Immunofect MG beads, indicates that transient holes of 5.4nm in diameter or larger are formed during immunoporation. The efficiencies of transfection of lymphocytes transfected with vectors coding for EGFP and lacZ were found to be within the range of 15-30% with high levels of cell viability of more than 90%. In addition, it was observed that mononuclear cells stimulated with PHA expressed transfected reporter genes with a higher efficiency. In conclusion, these results demonstrate that immunoporation using Immunofect MG beads can be used for the efficient transfection of primary lymphocytes with DNA or other macromolecules.

Factors that affect the efficiency of cell transfection by immunoporation.

Immunoporation is a recently discovered method that is able to transfect various human cell lines efficiently by targeting the cell surface antigens with antibody-coated beads. For this particular study, HL60, a cell line difficult to transfect by other methods, was used as a model to define the various parameters of the cell membrane that determine the efficiency of this method. The level of antigen expression on the cell surface was the first parameter to be analyzed and experiments indicated that there is a close correlation between the level of expression of surface antigens and the efficiency of immunoporation. The mixing speed, the bead to cell ratio, and the mixing time were all found to affect the ability of the antigen-coated beads to pull holes in the cells and it was found that for HL60 cells the optimum mixing speed was 40 rpm and the bead to cell ratio was 20:1 using a mixing time of 6 h.

Ego-integrity versus ego-despair: the effect of "accepting the past" on depression in older women.

The relative strength of the later-life personality process of ego-integrity, as operationalized by "accepting the past," was tested as a predictor of depression in a multivariate model containing other well established predictors-age, social support, physical dependency, and positive and negative affectivity. Seventy-three older Australian women living in supported accommodation completed an anonymous, self-report questionnaire. The results showed that "accepting the past" was a significant predictor in the multivariate model, along with social support, physical dependency, and positive affectivity. The results are discussed in terms of the utility of investigating later-life personality processes as potential interventions for alleviating depression in older people.

Effect of osmolarity and presence of serum on the efficiency of cell transfection using immunoporation.

Immunoporation is a novel method of cell transfection based upon the use of a new type of beads, Immunofect beads, that can be targeted to make holes in different types of cells depending on the type of bead used. It is known that the efficiency of transfection of cells by some techniques can be affected by the presence of serum and another important factor that appears to affect transfection efficiency and cell viability is the osmolarity of the transfection medium. This report presents studies on the effects of serum and varying osmolarity on the efficiency of transfection using immunoporation. The results clearly indicate that in hypertonic media the presence of serum decreases the efficiency of transfection. In the case of osmolarity, increasing the osmolarity of the immunoporation medium increases the efficiency of transfection but above about 650 mOsm this increasing efficiency is offset by the much lower viability of the cells.

An investigation into the suitability of silica beads for cell separations based on density perturbation.

This study has investigated possible alternative types of beads for fractionating cells on the basis of density perturbation. It is well known that uniform magnetic beads can be extremely important tools for separating cells by both magnetic separation techniques and density perturbation. However, because of the inherent expense associated with the use of magnetic beads, it was decided to study the possible use of inexpensive silica beads for density perturbation in terms of their attachment and modification of density of cells and to compare them with uniform Dynabeads. Silica beads were analyzed to determine their size and effect on the density of cells. Differentiated HL60 cells were used as a model system. As differentiation occurs, different levels of antigens are expressed on the cell surface and this results in different numbers of beads binding to cells. DMSO-differentiated HL60 cells were mixed with anti-CD11b-coated beads at a ratio of 20:1 (beads/cell), and gentle mixing was carried out at 20 degrees C on the end-over-end mixer. The binding of antibody-coated silica beads and Dynabeads to partially differentiated HL60 cells were compared. The conclusions reached on the basis of these experiments are that antibody-coated silica beads (Ab-coated silica) can be used as alternative beads for some cell fractionations. However, compared with Dynabeads, there are more beads that are only transiently associated with cells, possibly indicating that higher levels of detachment of beads from cells occur when silica beads are used. In addition, silica beads are usually heterogeneous in size and this would make it difficult to use these beads for the isolation of purified subpopulations of differentiated cells.

Fractionation of differentiating cells using density perturbation.

This paper describes the development of a new method for the fractionation of purified subpopulations of partially differentiated cells on continuous isopycnic gradients, using a density perturbation method based on the ability of cells to bind dense antibody-coated beads. Until now none of the available fractionation techniques, such as magnetic cell fractionation has been efficient for separating subpopulations of partially differentiated cells. The fractionation experiments described in this report used promyelocytic HL-60 and DMSO-induced granulocytic HL-60 cells as a model system. Populations of cells, modified by the binding of dense beads were fractionated on isotonic, isopycnic Optiprep gradients by centrifugation at 220xg for 90 min at 20 degrees C. Examination of the different gradient fractions showed that, as cells bind increasing numbers of beads, they are found in the denser regions of the isopycnic gradients. Indirect immunofluorescence was combined with flow cytometric techniques to characterise the fractionation of partially differentiated cells. Flow cytometric results confirmed that as antigenic determinants appear on the surface at higher levels of expression, the number of beads binding to each cell increased. Furthermore, after fractionation, when the bead-bound and non-bead-bound cells were cultured in the presence of DMSO, those cells that had bound more beads targeted to differentiated cells were found to achieve terminal differentiation faster than those cells that had not been associated with any beads.

Childbirth confidence: validating the Childbirth Self-Efficacy Inventory (CBSEI) in an Australian sample.

This study examined the reliability and validity of the Childbirth Self-Efficacy Inventory (CBSEI) (Lowe 1993) in a sample of 100 Australian women. Consistent with US data, the measure was shown to have high internal consistency. Validity of the instrument was determined by applying self-efficacy theory (Bandura 1982), which predicts that parity should have the largest effect on childbirth self-efficacy, followed by knowledge, then support and finally anxiety. Results revealed that having prior good birth experience and knowledge about childbirth had significant effects on childbirth self-efficacy. A factor analysis was performed to determine whether the original factor structure of this instrument held for Australian women. While outcome expectancies and self-efficacy expectancies emerged as distinct factors, the results showed that Australian women did not differentiate between active and second-stage labour. Rather than the two stages of labour emerging as dimensions of the CBSEI, two externally focused coping strategies were revealed.

An investigation into the possible use of hydrocyclones for the removal of yeast from beer.

This investigation has focused on the importance of hydrocyclone design and running conditions for the removal of yeast from suspensions. Experiments have been carried out using both a model system of yeast grown in suspension cultures and rough beer. It appeared that the most important factor was the design of the hydrocyclone with a positive correlation between higher residence times and acceleration with the efficiency of separation. The other factor that affected the separation efficiency was the concentration of the yeast suspension and a new phenomenon of a lower efficiency of hydrocyclones at very low feed concentrations has been observed. The separation efficiency was also affected by temperature with lower temperatures giving poorer separations.

A simple immunomagnetic bead-based technique for the detection of surface molecules capable of inducing T cell functional polarisation.

Activated T cells can release lymphokines selectively towards the site of contact with the target cell. In this way the specificity of the target-effector cell interaction can be maintained in spite of signalling being mediated by soluble factors (Mosmann, 1988, Immunol. Today 9, 306). However, this polarised phenotype is not expressed in resting T cells; rather it appears to be induced in the first minutes following T cell activation. In order to analyse single molecules for their ability to induce T cell polarisation, we devised a technique based on targeting different T cell surface molecules with specific antibodies immobilised on to immunomagnetic beads. The polarised phenotype was determined from observation of the microtubule organising centre being oriented towards the site of interaction with the bead. When applied to T cell lines, the technique permitted the classification of CD3 as a polarisation-inducing molecule, while no polarisation was found when targeting CD2, CD6 and CD8 molecules. This technique has a number of potential applications since it can, in principle, be applied to any cell surface molecule or cell type. Technical details and the sensitivity of the procedure are discussed.