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.

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.

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.

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.

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.

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.

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.

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.

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 epitope recognised by inhibitory monoclonal antibodies that react with a 51 kilodalton merozoite surface antigen in Plasmodium falciparum.

Monoclonal antibodies designated 8G10/48 and 9E3/48 raised against mature asexual blood stages of Plasmodium falciparum inhibit parasite growth in vitro. Both antibodies bind to an epitope which includes the linear sequence Ser Thr Asn Ser and which is present in a cDNA clone from a P. falciparum expression library. These antibodies recognise a glycosylated antigen of approximately 51 kDa which is located on the merozoite surface membrane.

The effects of haloperidol upon temporal information processing by patients with Tourette's syndrome.

Tourette's syndrome patients treated successfully with haloperidol, untreated patients, and healthy controls were studied with tests of temporal discrimination and measures of transmitted information shown previously to be sensitive to brain dysfunction. Untreated patients showed no impairment of temporal processing while those treated with haloperidol showed significant deficit in amount of transmitted information comparable to prior studies of brain syndromes.

Effects of trifluoperazine, chlorpromazine, and haloperidol upon temporal information processing by schizophrenic patients.

Healthy controls, unmedicated, actively symptomatic schizophrenics, and similar patients undergoing treatment with either trifluoperazine, chlorpromazine, or haloperidol were studied with tests of temporal discrimination and measures of transmitted information shown previously to be sensitive to various kinds of brain dysfunction, including haloperidol effects in a nonpsychotic population. Variations in psychophysical method, cognitive load, discrimination complexity, and sense-mode conditions permitted representative sampling of the temporal processing. Untreated, actively psychotic patients showed no impairment of temporal processing while all three antipsychotic medications were associated with significant deficit; trifluoperazine and haloperidol produced the most deficit, with chlorpromazine in the middle between the higher potency drugs on the one hand and unmedicated patients and healthy controls on the other.

Temporal information processing by alcoholics.

In time-discrimination tests using visual and auditory stimuli, more information was transmitted by social drinkers than by alcoholics, and cognitively unimpaired alcoholics transmitted more information than did cognitively impaired alcoholics.

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.

Signal pulse-rate and judged duration.

Therories which construct perception of time from content of input predict monotonic functions of rate-judged duration of stimuli, and do not account for intersensory differences. Two experiments required Ss to compare directly the durations of paird lights or sounds pulsed at various rates to produce discriminable beats and flickers (6.0, 10.0, 14.0 Hz), and steady signals. Pulsed lights, not sounds, were judged longer than steady, and this visual effect was identical for all flicker rates. Faster pulsled sounds and lights were judged longer than slower ones for all frequency combinations except for 10.0 to 14.0-Hz comparisons with vision. No monotonic function of ratejudged durations of pulses was obtained; the effect was all-or-none. Although pulse rate did affect judged duration, neither simple functions nor symmetry across senses was found.

Temporal information processing and psychopathology.

Prior studies showed impaired temporal discrimination by schizophrenic and neurologic patients reflected in decreased information transmission. This report describes a study of 8 more carefully diagnosed schizophrenic patients, separating those with neurologic signs. Using temporal discrimination tasks involving two psychophysical methods, 8 schizophrenic patients with no organic signs did not differ from 17 nonpsychotic, nonorganic patients; and organic group (n = 5) transmitted less information than the other patient samples. It is suggested that prior results were a product of casual diagnosis that ignored organic factors; reduced efficiency of temporal processing is associated predominantly with neurologic impairment.