Early psychosis research at Orygen, The National Centre of Excellence in Youth Mental Health.

BACKGROUND: Specialised early intervention (SEI) programs have offered individuals with psychotic disorders and their families new hope for improving illness trajectories and outcomes. The Early Psychosis Prevention and Intervention Centre (EPPIC) was one of the first SEI programs developed in the world, providing services for young people experiencing their first episode of psychosis. METHODS: We conducted a narrative synthesis of controlled and uncontrolled studies that have been conducted at EPPIC. DISCUSSION: The history of the EPPIC model is first described. This is followed by a discussion of clinical research emerging from EPPIC, including psychopharmacological, psychotherapeutic trials and outcome studies. Neurobiological studies are also described. Issues pertaining to the conduct of clinical research and future research directions are then described. Finally, the impact of the EPPIC model on the Australian environment is discussed.

Immunogenicity and safety of the 9-valent HPV vaccine in men.

OBJECTIVES: This study was designed to evaluate the immunogenicity and tolerability of a prophylactic 9-valent HPV (types 6/11/16/18/31/33/45/52/58) VLP (9vHPV) vaccine in young men 16-26 years of age in comparison to young women 16-26 years of age (the population that was used to establish 9vHPV vaccine efficacy). Safety and immunogenicity data from this study will be used to bridge 9vHPV vaccine efficacy findings in 16-26 year old women to 16-26 year old men. METHODS: This study enrolled 1106 heterosexual men (HM) and 1101 women who had not yet received HPV vaccination. In addition, 313 men having sex with men (MSM) were enrolled and were evaluated separately for immunogenicity because previous results showed that antibody responses to quadrivalent HPV (types 6/11/16/18) VLP (qHPV) vaccine were lower in MSM than in HM. All subjects were administered a 3-dose regimen (Day 1, Month 2, Month 6) of 9vHPV vaccine. Serum samples were collected for anti-HPV assays. Safety information was collected for ∼ 12 months. RESULTS: The geometric mean titers (GMTs) for HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58 for HM were non-inferior to those of women at Month 7. For all vaccine HPV types, Month 7 GMTs were numerically lower in MSM than in HM. Over 99.5% of subjects were seropositive at Month 7 for each vaccine HPV type. Administration of 9vHPV vaccine to both 16-26 year old men and women was generally well tolerated. CONCLUSIONS: These results support bridging the efficacy findings with 9vHPV vaccine in young women 16-26 years of age to men 16-26 years of age.

An investigation into the enhancement of sea-spray exposed fingerprints on glass.

Fingerprints are considered one of the best forms of personal identification. While numerous enhancement techniques exist to develop fingerprints under various conditions, the enhancement of fingerprints exposed to sea spray aerosol (SSA) still remains problematic. 1056 fingerprints from four donors, using a depletion series and triplicate repeats, were deposited onto glass panels and exposed to SSA for 1 week and 1 month. Control prints were deposited in the same manner and left under laboratory conditions. All prints were enhanced using fingerprint enhancement techniques available to Forensic Police Officers and subsequently examined for identifiability by a Fingerprint Expert. Significantly fewer identifiable prints (p<0.01) were developed after exposure to SSA for 1 month (11%) compared to exposure for 1 week (69%) (compared to the control prints 99%) for all enhancement techniques. After 1 week's exposure, all techniques enhanced over 50% of prints, except SPR white (12%), with iron (III) oxide and Wetwop™ white producing over 90% identifiable prints. Only iron (III) oxide, Wetwop™ white and SPR black returned any identifiable prints following 1 month's SSA exposure. Iron (III) oxide being significantly better (p<0.01, 67%) than the other techniques. Iron (III) oxide suspension and Wetwop™ white were found to be superior at enhancing prints at both SSA exposure times.

The fear factor: drivers and barriers to follow-up screening for human papillomavirus-related anal cancer in men who have sex with men.

Human papillomavirus (HPV)-related anal cancer incidence is rising in men who have sex with men (MSM). Effective screening strategies exist, but many patients are lost to follow-up (LTF). We studied factors impacting screening compliance to recommended annual screening visits. Retrospective chart review identified MSM with anal dysplasia. MSM were grouped as regular screeners (regular to follow-up [RF]) (≥1 visit/year), lost to follow-up (LTF) (>1 year since previous screening) and LTF who then returned for screening (lost came back [LCB]). From June 2007 to March 2008, subjects completed a questionnaire in-person at the time of screening or via telephone (LTF). Questionnaires were completed after anal dysplasia diagnosis. One hundred and ninety-five MSM were enrolled (96 RF, 50 LTF and 49 LCB). RF were compliant for 4.8 years; LTF were lost for 2.3 years. LCB were previously lost for 5.6 years before returning. Mean knowledge score of screening procedures was larger in RF versus LTF (P < 0.001). MSM with more sexual partners in the past six months were more likely to be LCB versus LTF (P = 0.05). RF were more likely to describe their HPV diagnosis as 'upsetting' (P = 0.003). RF were more likely driven by physical symptoms versus LTF (P = 0.002). MSM with high-grade intraepithelial lesions (HSIL) were more likely to be RF versus those with low-grade intraepithelial lesions (P = 0.001. Positive predictors for screening compliance include an upsetting experience during the HPV diagnosis, physical symptoms driving the initial visit and HSIL. Engaging patients in a firm, salient approach may facilitate follow-up compliance.

High prevalence of anal squamous intraepithelial lesions and squamous-cell carcinoma in men who have sex with men as seen in a surgical practice.

INTRODUCTION: Anal high-grade squamous intraepithelial lesions are probable invasive anal squamous-cell cancer precursors, and although unproved, treatment of high-grade squamous intraepithelial lesions may prevent progression to anal squamous-cell cancer. Men who have sex with men are often treated for benign anorectal disorders without consideration given to the possibility of concurrent high-grade squamous intraepithelial lesions or anal squamous-cell cancer. We determined the prevalence of anal high-grade squamous intraepithelial lesions and anal squamous-cell cancer in an urban surgical practice of men who have sex with men referred for treatment of anal condyloma and other benign noncondylomatous anal disorders. METHODS: One hundred thirty-one HIV-positive and 69 HIV-negative men who have sex with men referred for surgical treatment of presumed benign anorectal disease were evaluated by anal cytology, high-resolution anoscopy, and biopsy. Anal cytology and histology were reported with a modified Bethesda classification. RESULTS: One hundred fifty-seven patients (79 percent) were referred for condyloma, 4 (2 percent) for anal squamous intraepithelial lesions (anal high-grade squamous intraepithelial lesions) diagnosed by primary care providers, and 39 (19 percent) for other benign anorectal disorders. One hundred forty-three patients (93 percent) had abnormal anal cytology, with 107 (54 percent) having high-grade squamous intraepithelial lesions on cytology. Biopsy results revealed 120 patients (60.0 percent) with high-grade squamous intraepithelial lesions and 5 patients (3 percent) with invasive squamous-cell carcinoma. Four of five men with anal squamous-cell cancer were HIV positive. Fourteen men (36 percent) who have sex with men referred for noncondylomatous benign anal disorders had high-grade squamous intraepithelial lesions, and three (8 percent) had anal squamous-cell cancer. High-grade squamous intraepithelial lesions and anal squamous-cell cancer were seen most often at the squamocolumnar junction. CONCLUSIONS: Men who have sex with men referred for treatment of either condyloma or noncondylomatous benign anorectal disease had a high prevalence of anal high-grade squamous intraepithelial lesions and anal squamous-cell cancer. All men who have sex with men referred for treatment of benign anorectal disease should have high-resolution anoscopy and aggressive biopsy of all abnormal areas. Treatment of external lesions alone could miss high-grade squamous intraepithelial lesions or anal squamous-cell cancer.

Cdc25-dependent activation of cyclin A/cdk2 is blocked in G2 phase arrested cells independently of ATM/ATR.

Cyclin A/cdk2 is active during S and G2 phases of the cell cycle, but its regulation and function during G2 phase is poorly understood. In this study we have examined the regulation of cyclin A/cdk2 activity during normal G2 phase progression and in genotoxin-induced G2 arrest. We show that cyclin A/cdk2 is activated in early G2 phase by a cdc25 activity. In the G2 phase checkpoint arrest initiated in response to various forms of DNA damage, the cdc25-dependent activation of both cyclin A/cdk2 and cyclin B1/cdc2 is blocked. Ectopic expression of cdc25B, but not cdc25C, in G2 phase arrested cells efficiently activated both cyclin A/cdk2 and cyclin B1/cdc2. Finally, we demonstrate that the block in cyclin A/cdk2 activation in the G2 checkpoint arrest is independent of ATM/ATR. We speculate that the ATM/ATR-independent block in G2 phase cyclin A/cdk2 activation may act as a further layer of checkpoint control, and that blocking G2 phase cyclin A/cdk2 activation contributes to the G2 phase checkpoint arrest.

Forkhead transcription factors, Fkh1p and Fkh2p, collaborate with Mcm1p to control transcription required for M-phase.

BACKGROUND: The 'CLB2 cluster' in Saccharomyces cerevisiae consists of approximately 33 genes whose transcription peaks in late G2/early M phase of the cell cycle. Many of these genes are required for execution of the mitotic program and then for cytokinesis. The transcription factor SFF (SWI5 factor) is thought to regulate a program of mitotic transcription in conjunction with the general transcription factor Mcm1p. The identity of SFF has yet to be determined; hence further understanding of the mechanisms that regulate entry to M phase at the transcriptional level requires characterization of SFF at the molecular level. RESULTS: We have purified the biochemical activity corresponding to SFF and identified it as the forkhead transcription factor Fkh2p. Fkh2p assembles into ternary complexes with Mcm1p on both the SWI5 and CLB2 cell-cycle-regulated upstream activating sequence (UAS) elements in vitro, and in an Mcm1 p-dependent manner in vivo. Another closely related forkhead protein, Fkh1p, is also recruited to the CLB2 promoter in vivo. We show that both FKH1 and FKH2 play essential roles in the activation of the CLB2 cluster genes during G2-M and in establishing their transcriptional periodicity. Hence, Fkh1p and Fkhp2 show the properties expected of SFF, both in vitro and in vivo. CONCLUSIONS: Forkhead transcription factors have redundant roles in the control of CLB2 cluster genes during the G2-M period of the cell cycle, in collaboration with Mcm1p.

Anal dysplasia in men who have sex with men.

Risk factors for anal cancer include anal intercourse and infection with multiple strains of human papillomavirus, the causative agent of anal precancerous dysplasia. Several recent studies have shown that HIV-seropositive gay men are at greater risk for anal dysplastic lesions than seronegative gay men. Moreover, the risk for detection and progression of dysplastic lesions grows as the CD4+ cell count declines. A surgeon with a practice that includes gay men referred for anorectal disease presents data regarding the high prevalence of anal dysplasia in his patients.

Ethnicity study and non-selective screening for haemoglobinopathies in the antenatal population of central Manchester.

The objective of this study was to determine the frequency of ethnic groups within the antenatal population in central Manchester and thereby ensure that the haemoglobinopathy service was targeting the correct population and their needs. Ethnic data collection records of 6718 patients were analysed over a 7 month period. Of these 62.3% stated that they were White, 13.2% Asian, 7.9% Black, 3.8% Chinese or 'other ethnic groups' and 12.7% gave no information about their ethnic background. A subset of 1144 patients were screened for haemoglobinopathies over a 1 month period. The incidence of haemoglobinopathies within the screened population was 2.62%, and comprised 0.69% beta thalassaemia trait, 1.22% sickle cell trait, 0.43% haemoglobin C trait and 0.26% haemoglobin D trait. The total incidence of haemoglobinopathies was highest within the Black population (18.2%), followed by the no information group (5.6%), Asian (3.35%) and white (0.26%). The high proportion of ethnic minorities and the significant carrier frequency in the no information group, support our view that non-selective screening should be offered to the antenatal population of central Manchester.

An evaluation of the Bio-Rad Variant Haemoglobin Testing System for the detection of haemoglobinopathies.

The Bio-Rad Variant Haemoglobin Testing System is an automated analyser which uses the principle of cation exchange high performance liquid chromatography. This evaluation was undertaken to examine the effectiveness of the instrument as a screening mechanism to assist in the diagnosis of haemoglobinopathies. The ability to quantify haemoglobins A2 and F and to 'flag' other haemoglobin variants was tested. Within-batch precision was excellent and between-batch precision was good. Linearity and sensitivity compared favourably with the manufacturer's published ranges. The level of carry-over for haemoglobins F, S and A was less than 0.25%. The mean carry-over for haemoglobin A2 was 2.08%. This higher figure reflected the smaller absolute difference between the high and low samples for this parameter. The instrument never failed to indicate the presence of an abnormal haemoglobin in 271 selected samples. The instrument was reliable throughout the evaluation and at no time was a run aborted.

Redox regulation of the mitogen-activated protein kinase pathway during lymphocyte activation.

We have previously demonstrated an obligatory requirement for intracellular reactive oxygen species generation during T lymphocyte activation, and have proposed that intracellular reactive oxygen species may act as signalling agents in the regulation of certain cellular processes, for example, during cell cycle entry. To test this hypothesis, we have been interested to determine which, if any, cell cycle entry events are affected by oxidative signalling. In earlier studies, we have identified the transcription factors NF-kappa B and AP-1 as molecular targets for oxidative signalling processes during cell cycle entry, and have shown that oxidative signalling is involved in the regulation of early changes in gene expression during the G0 to G1 phase transition. To extend these initial observations, we have examined the effect of antioxidant treatment on the activity of the mitogen-activated protein kinases erk1 and erk2, as members of a signal transduction pathway known to directly regulate transcription factor function. Using as a probe cysteamine, an aminothiol compound with both antioxidant and antiproliferative activity, we have identified erk2, a key element of the MAP kinase pathway, as being responsive to oxidative signalling during lymphocyte activation. These observations provide further evidence to suggest a role for intracellular oxidant generation as a regulatory mechanism during cell cycle entry, and establish a link between oxidative signalling and other aspects of the intracellular signalling network that is activated in response to mitogenic stimulation.

Oxidative signalling and gene expression during lymphocyte activation.

We previously have demonstrated an obligatory requirement for intracellular reactive oxygen species (ROS) generation during T lymphocyte activation, and have proposed that ROS may act as signalling agents in the regulation of certain cellular processes, for example, during cell cycle entry. In order to test this hypothesis, we have been interested to determine which, if any, cell cycle entry events are affected by oxidative signalling. Given the requirement for both oxidative signalling and altered gene expression during the G0 to G1 phase transition, we have attempted to establish the extent to which oxidative signalling affects global gene expression patterns during cell cycle entry, and to isolate and characterize mRNAs whose expression patterns are responsive to oxidative signalling during this process. Using differential display in a phenotypic screening approach, we have identified 10 mRNA species whose expression patterns were altered in response to inhibition of oxidative signalling during cell cycle entry. The expression patterns of 4 of these 10 mRNAs were unaffected during cell cycle arrest caused by a different mechanism, cyclosporin A-induced interference with calcineurin-mediated signalling events, implying that the altered expression patterns seen were not simply a consequence of cell cycle arrest. This suggests that the expression of these 4 mRNAs is regulated by a mechanism both necessary for cell cycle entry and sensitive to oxidative signalling. RNAse protection assays confirmed that 2 of these 4 mRNAs were indeed responsive to redox regulation. These observations strongly suggest an involvement for oxidative signalling in the regulation of gene expression during the G0 to G1 phase transition, in peripheral blood mononuclear cells at least.

Transcription factors as targets for oxidative signalling during lymphocyte activation.

We previously have demonstrated a requirement for oxidative events during cell cycle entry in T lymphocytes and have hypothesised that reactive oxygen species may act as intracellular signalling agents during lymphocyte activation. In the current study, cysteamine, an aminothiol compound with antioxidant activity, has been used to further investigate the role of oxidative signalling during lymphocyte activation. Treatment of normal human peripheral blood lymphocytes with cysteamine in vitro was found to inhibit proliferation in a dose-dependent manner, with essentially complete inhibition occurring at a dose of 400 microM. This inhibitory effect was limited to the first 2 h after mitogenic activation, localizing the time-frame of action of cysteamine to within the commitment period. It therefore was of interest to establish which, if any, commitment events were affected by oxidative signalling during cell cycle entry. Taking the IL-2 gene as a candidate, we examined the effect of cysteamine treatment on early gene expression during lymphocyte activation, and on the activity of transcription factors AP-1, NF-kappa B, NF-AT and Oct1, whose functions are required for expression of the IL-2 mRNA. Cysteamine treatment inhibited both expression of the IL-2 mRNA and secretion of IL-2 into the culture medium. The inhibitory effect of cysteamine may be mediated at least in part by an effect on transcription factor function, as the DNA binding activities of AP-1 and NF-kappa B extracted from mitogen-stimulated cells were significantly inhibited by cysteamine treatment. Interestingly, Oct1 and NF-AT DNA binding activity were not affected by cysteamine treatment, suggesting that oxidative signalling processes operate in a selective manner. The identification of regulatory proteins, such as transcription factors, as molecular targets for oxidative signalling provides further evidence to implicate oxidative signalling as being intimately involved in the G0 to G1 phase transition in T lymphocytes.

Prolonged expression of c-jun and associated activity of the transcription factor AP-1, during apoptosis in a human leukaemic cell line.

The product of the c-jun gene is a component of the transcription factor AP-1, which plays a critical regulatory role in the cellular response to certain proliferative stimuli. In this report, we demonstrate the prolonged expression of c-jun mRNA during apoptosis induced in a human leukaemic T-cell line, CEM C7, by treatment with either dexamethasone or gamma-radiation. However, overexpression of the c-jun mRNA was not accompanied by either increased expression of jun protein, or increased AP-1 DNA binding activity. Indeed, a decrease in AP-1 DNA binding activity was seen in response to both inducing stimuli. This decrease in AP-1 binding activity may be mediated by an increase in the activity of an AP-1 inhibitory factor, as the steady state levels of jun protein remained constant during the onset of apoptosis, and cytosolic AP-1 inhibitory activity was found to increase, concomitant with the decrease in AP-1 DNA binding activity. Our data demonstrate the complexity of the mechanisms involved in the regulation of c-jun expression during the onset of apoptosis, and suggest a novel mode for the regulation of AP-1 activity during the cessation of proliferation.

Localization of IFN-gamma and IL-6 mRNA in murine intestine by in situ hybridization.

Previous studies have highlighted the importance of CD4+ T cells in regulation of IgA responses and have indicated a functional heterogeneity among these cells between inductive (Peyer's patch) and effector (lamina propria) sites in the intestine. To determine whether these functional differences could be accounted for by differences in cytokine profile of cells in each of these sites, the distribution of mRNA for interferon-gamma (IFN-gamma) and interleukin-6 (IL-6) was investigated by in situ hybridization techniques using 35S-labelled riboprobes. Whereas message for IL-6 is abundant in all regions of the lamina propria from the base of the mucosa to the tips of the villi, very little is expressed in Peyer's patches or in the epithelium. In contrast, message for IFN-gamma is expressed predominantly by cells localized only in the base of the lamina propria and, as with IL-6, very little message was detected in Peyer's patches although occasional strongly positive IFN-gamma cells were observed in the epithelium. These results indicate that, at least in the absence of deliberate intestinal stimulation, functional expression of these cytokines is restricted to effector rather than induction sites in the intestine. This is consistent with our previous observations demonstrating a requirement for T-cell signals in promoting post-extravasation differentiation and proliferation of IgA-committed B cells in vivo and the implications of these findings to the role of the Th1 and Th2 subsets of CD4+ cells in mucosal immune responses is discussed.

Isolation of a cDNA clone, encoding the ribosomal protein S20, downregulated during the onset of apoptosis in a human leukaemic cell line.

In this report we describe the isolation, by differential screening, and characterization of a cDNA clone downregulated fivefold in association with the induction of apoptosis in the human leukaemic cell line CEM C7. DNA sequence analysis identified this clone as representing the gene encoding the S20 ribosomal protein. Interestingly, the expression of the S20 mRNA was downregulated early during the induction of apoptosis in this model system, prior to the onset of DNA fragmentation and other morphological changes associated with cell death, suggesting some degree of involvement of this particular gene in the biochemical events that occur during the onset of cell death.

Isolation of a cDNA clone, encoding a human beta-galactoside binding protein, overexpressed during glucocorticoid-induced cell death.

In this report we describe the isolation and characterization of a cDNA clone overexpressed tenfold during the induction of apoptosis in the glucocorticoid-sensitive human leukaemia cell line CEM C7. This clone was shown by DNA sequence analysis to represent the human HL14 gene, encoding a beta-galactoside binding protein, the mouse homologue of which has recently been reported to act as a cell growth inhibitory factor.