Chemo-enzymatic synthesis of unnatural amino acids

A general chemo-enzymatic process has been developed to prepare enantiomerically pure L- and D-amino acids in high yield by deracemisation of racemic starting materials. The method has been developed from initial academic studies to be a robust, scalable industrial process. Unnatural amino acids, in high optical purity, are a rapidly growing class of intermediates required for pharmaceuticals, agrochemicals and other fine chemical applications. However, no single method has proven sufficiently adaptable to prepare these compounds generally at large scale. Our approach uses an enantioselective oxidase biocatalyst and a non-selective chemical reducing agent to effect the stereoinversion of one enantiomer and can result in an enantiomeric excess of > 99 % from a starting racemate, and product yields over 90 %. The current approach compares very favourably to resolution methods which have a maximum single pass yield of 50 %. Efficient methods have been developed to adapt the biocatalyst used in this process towards new target compounds and to optimise key factors which improve the process efficiency and offer competitive economics at scale.

Exploring the factor structure of a new self- and other-oriented alcohol expectancy scale relating to sexual coercion and vulnerability

Introduction and Aim: Sexual assaults commonly involve alcohol use by the perpetrator, victim, or both. Beliefs about alcohol’s effects may impact on people’s perceptions of and responses to men and women who have had such experiences while intoxicated from alcohol. This study aimed to develop an alcohol expectancy scale that captures young adults’ beliefs about alcohol’s role in sexual aggression and victimisation. Design and Methods: Based on pilot focus groups, an initial pool of 135 alcohol expectancy items was developed, checked for readability and face validity, and administered via a cross-sectional survey to 201 male and female university students (18-25 years). Items were specified in terms of three target drinkers: self, men, and women. In addition, a social desirability measure was included. Results: Principal Axis Factoring revealed a 4-factor solution for the targets men and women and a 5-factor solution for the target self with 72 items retained. Factors related to sexual coercion, sexual vulnerability, confidence, self-centredness, and negative cognitive and behavioural effects. Social desirability issues were evident for the target self, but not for the targets men and women. Discussion and Conclusions: Young adults link alcohol’s effects with sexual vulnerabilities via perceived risky cognitions and behaviours. Due to social desirability, these expectancies may be difficult to explicate for the self but may be accessible instead via other-oriented assessment. The Sexual Coercion and Vulnerability Alcohol Expectancy Scale has potential as a tool to elucidate the established tendency for observers to excuse intoxicated sexual perpetrators while blaming intoxicated victims.

Study Protocol - Alcohol Management Plans (AMPs) in remote indigenous communities in Queensland: their impacts on injury, violence, health and social indicators and their cost-effectiveness

Background In 2002/03 the Queensland Government responded to high rates of alcohol-related harm in discrete Indigenous communities by implementing alcohol management plans (AMPs), designed to include supply and harm reduction and treatment measures. Tighter alcohol supply and carriage restrictions followed in 2008 following indications of reductions in violence and injury. Despite the plans being in place for over a decade, no comprehensive independent review has assessed to what level the designed aims were achieved and what effect the plans have had on Indigenous community residents and service providers. This study will describe the long-term impacts on important health, economic and social outcomes of Queensland’s AMPs. Methods/Design The project has two main studies, 1) outcome evaluation using de-identified epidemiological data on injury, violence and other health and social indicators for across Queensland, including de-identified databases compiled from relevant routinely-available administrative data sets, and 2) a process evaluation to map the nature, timing and content of intervention components targeting alcohol. Process evaluation will also be used to assess the fidelity with which the designed intervention components have been implemented, their uptake and community responses to them and their perceived impacts on alcohol supply and consumption, injury, violence and community health. Interviews and focus groups with Indigenous residents and service providers will be used. The study will be conducted in all 24 of Queensland’s Indigenous communities affected by alcohol management plans. Discussion This evaluation will report on the impacts of the original aims for AMPs, what impact they have had on Indigenous residents and service providers. A central outcome will be the establishment of relevant databases describing the parameters of the changes seen. This will permit comprehensive and rigorous surveillance systems to be put in place and provided to communities empowering them with the best credible evidence to judge future policy and program requirements for themselves. The project will inform impending alcohol policy and program adjustments in Queensland and other Australian jurisdictions. The project has been approved by the James Cook University Human Research Ethics Committee (approval number H4967 & H5241).

The alcohol culture of mature-aged women : similarities and differences across borders

Introduction: Relatively few attempts have been made to describe and understand women’s alcohol consumption beyond adolescence and young adulthood. In particular, there has been a lack of studies focusing on the alcohol culture that surrounds and guides mature-aged women’s drinking. As part of a larger cross-national comparison, the present study sought to address this gap by identifying the shared beliefs and values that impact on drinking outcomes among mature-aged women in Sweden and Australia. Method: The study was guided by an ethnographic methodology. To generate data, a series of semi-structured interview were conducted with 17 Australian (age = 45-57 years; M = 52.1, SD = 3.9) and 19 Swedish (age = 45-58 years; M = 52.2, SD = 4.8) women. All interviews were transcribed verbatim and thematically analysed. Results: With age, the focus of alcohol as a single purpose vehicle for intoxication had given way to a focus on the enjoyment and ritual of drinking itself; taste had become increasingly important and alcohol was strongly associated with pleasurable environments and experiences. The view of alcohol as a taste experience was particularly pronounced among the Swedish women, with alcohol (most commonly wine) often seen as inseparable from food. Among the Swedish women, this view of alcohol was also associated with a strong de-emphasis of the pharmacological effects of alcohol. In contrast, several Australian women understood and used alcohol as relief for anxiety and stress. Moderate drinking was linked to the social construction of both age and gender in the two samples, while heavy or abusive alcohol consumption was associated with strong proscriptive norms and stigma. Conclusions: Australian and Swedish women share a number of beliefs and values around alcohol, however, these findings also show unique country-level differences. Implications for drinking outcomes are discussed.

The Queensland Indigenous alcohol diversion program : a health agency’s journey

Drug and alcohol diversion programs provide offenders with the opportunity to divert from the criminal justice or child safety systems, and enter into treatment to address their illicit drug or alcohol use. However, low participation by Indigenous Australians in diversion programs has been recognised as an issue, with Indigenous Australians being much less likely to be diverted into treatment (NIDAC 2009: 9). QIADP represents a unique opportunity to improve Indigenous access to diversion programs. QIADP is an Indigenous-specific alcohol diversion program in its final, third year as a pilot, with the evaluation due December 2009. Many lessons have been learnt by Queensland Health as to what works and doesn’t work in the provision of alcohol-related treatment with this population, including how partnerships with other governmental departments and NGOs can enhance the quality of treatment and ways to build clinical cultural competence in the workforce and programmatic system. This presentation shares the practical lessons QH has learnt in delivering alcohol treatment within an Indigenous-specific diversion program. This includes solutions that others may find useful for application elsewhere, such as the holistic range of treatment options found helpful, and the relationship issues to work through to support a partnership response.

Failure of amino acid homeostasis causes cell death following proteasome inhibition

The ubiquitin-proteasome system targets many cellular proteins for degradation and thereby controls most cellular processes. Although it is well established that proteasome inhibition is lethal, the underlying mechanism is unknown. Here, we show that proteasome inhibition results in a lethal amino acid shortage. In yeast, mammalian cells, and flies, the deleterious consequences of proteasome inhibition are rescued by amino acid supplementation. In all three systems, this rescuing effect occurs without noticeable changes in the levels of proteasome substrates. In mammalian cells, the amino acid scarcity resulting from proteasome inhibition is the signal that causes induction of both the integrated stress response and autophagy, in an unsuccessful attempt to replenish the pool of intracellular amino acids. These results reveal that cells can tolerate protein waste, but not the amino acid scarcity resulting from proteasome inhibition.

Mechanisms of mono- and poly-ubiquitination : ubiquitination specificity depends on compatibility between the E2 catalytic core and amino acid residues proximal to the lysine

Ubiquitination involves the attachment of ubiquitin to lysine residues on substrate proteins or itself, which can result in protein monoubiquitination or polyubiquitination. Ubiquitin attachment to different lysine residues can generate diverse substrate-ubiquitin structures, targeting proteins to different fates. The mechanisms of lysine selection are not well understood. Ubiquitination by the largest group of E3 ligases, the RING-family E3 s, is catalyzed through co-operation between the non-catalytic ubiquitin-ligase (E3) and the ubiquitin-conjugating enzyme (E2), where the RING E3 binds the substrate and the E2 catalyzes ubiquitin transfer. Previous studies suggest that ubiquitination sites are selected by E3-mediated positioning of the lysine toward the E2 active site. Ultimately, at a catalytic level, ubiquitination of lysine residues within the substrate or ubiquitin occurs by nucleophilic attack of the lysine residue on the thioester bond linking the E2 catalytic cysteine to ubiquitin. One of the best studied RING E3/ E2 complexes is the Skp1/Cul1/F box protein complex, SCFCdc4, and its cognate E2, Cdc34, which target the CDK inhibitor Sic1 for K48-linked polyubiquitination, leading to its proteasomal degradation. Our recent studies of this model system demonstrated that residues surrounding Sic1 lysines or lysine 48 in ubiquitin are critical for ubiquitination. This sequence-dependence is linked to evolutionarily conserved key residues in the catalytic region of Cdc34 and can determine if Sic1 is mono- or poly-ubiquitinated. Our studies indicate that amino acid determinants in the Cdc34 catalytic region and their compatibility to those surrounding acceptor lysine residues play important roles in lysine selection. This may represent a general mechanism in directing the mode of ubiquitination in E2 s.

BDNF SNPs are implicated in comorbid alcohol dependence in schizophrenia but not in alcohol-dependent patients without schizophrenia

Aims The functional BDNF single nucleotide polymorphism (SNP) rs6265 has been associated with many disorders including schizophrenia and alcohol dependence. However, studies have been inconsistent, reporting both positive and negative associations. Comorbid alcohol dependence has a high prevalence in schizophrenia so we investigated the role of rs6265 in alcohol dependence in Australian populations of schizophrenia and alcohol dependent patients. Methods Two BDNF SNPs rs6265 and a nearby SNP rs7103411 were genotyped in a total of 848 individuals. These included a schizophrenia group (n = 157) and a second schizophrenia replication group (n = 235), an alcohol dependent group (n = 231) that had no schizophrenia diagnosis and a group of healthy controls (n = 225). Results Allelic association between rs7103411 and comorbid alcohol dependence was identified (P = 0.044) in the primary schizophrenia sample. In the replication study, we were able to detect allelic associations between both BDNF SNPs and comorbid alcohol dependence (rs6265, P = 0.006; rs7103411, P = 0.014). Moreover, we detected association between both SNPs and risk-taking behaviour after drinking (rs6265, P = 0.005; rs7103411, P = 0.009) and we detected strong association between both SNPs and alcohol dependence in males (rs6265, P = 0.009; rs7103411, P = 0.013) while females showed association with multiple behavioural measures reflecting repetitive alcohol consumption. Haplotype analysis revealed the rs6265-rs7103411 A/C haplotype is associated with comorbid alcohol dependence (P = 0.002). When these SNPs were tested in the non-schizophrenia alcohol dependent group we were unable to detect association. Conclusion We conclude that these BDNF SNPs play a role in development of comorbid alcohol dependence in schizophrenia while our data does not indicate that they play a role in alcohol dependent patients who do not have schizophrenia.

A qualitative exploration of young Australian adults’ understanding of and explanations for alcohol-involved rape

Little is known about the beliefs that underlie the biased attributions that typically characterise people’s perceptions of intoxicated sexual perpetrators and their victims. Guided by consensual qualitative research, we explored young Australian adults’ (18-25 years; N = 15) attributions for an alcohol-involved rape based on focus groups and interviews. Prominent themes indicated that participants rarely labelled the assault as rape and, instead, adhered to miscommunication explanations. Participants emphasised the developmental value of the victimisation experience although recognising its harmful consequences. Both perpetrator and victim were held strongly responsible based on perceived opportunities to prevent the assault but implicit justifications were, nevertheless, evident. As such, explicit and implicit attributions were contradictory, with the latter reflecting the attributional double standard previously observed in quantitative rape-perception research. Findings underscore the need to challenge pervasive rape myths and equip young adults with knowledge on how to respond supportively to the commonly stigmatised victims of rape.

Effect of the increase in "alcopops" tax on alcohol-related harms in young people : a controlled interrupted time series

Objective: To measure alcohol-related harms to the health of young people presenting to emergency departments (EDs) of Gold Coast public hospitals before and after the increase in the federal government "alcopops" tax in 2008. Design, setting and participants: Interrupted time series analysis over 5 years (28 April 2005 to 27 April 2010) of 15-29-year-olds presenting to EDs with alcohol-related harms compared with presentations of selected control groups. Main outcome measures: Proportion of 15-29-year-olds presenting to EDs with alcohol-related harms compared with (i) 30-49-year-olds with alcohol-related harms, (ii)15-29-year-olds with asthma or appendicitis, and (iii) 15-29-yearolds with any non-alcohol and non-injury related ED presentation. Results: Over a third of 15-29-year-olds presented to ED with alcohol-related conditions, as opposed to around a quarter for all other age groups. There was no significant decrease in alcohol-related ED presentations of 15-29-year-olds compared with any of the control groups after the increase in the tax. We found similar results for males and females, narrow and broad definitions of alcoholrelated harms, under-19s, and visitors to and residents of the Gold Coast. Conclusions: The increase in the tax on al copops was not associated with any reduction in alcohol-related harms in this population in a unique tourist and holiday region. A more comprehensive approach to reducing alcohol harms in young people is needed.

Good choices, great future : an applied theatre prevention program to reduce alcohol-related risky behaviours during Schoolies

Introduction and Aims. The contextual and temporal factors of post-school celebratory events ('Schoolies') place young people at elevated risk of excessive drinking compared with other social occasions. This study investigates the impact of an applied theatre prevention program 'Choices' in reducing the risk of drinking and other risk behaviours during Schoolies celebrations. Design and Methods. Choices was delivered in the last term of Year 12 across 28 North Queensland schools. A total of 352 school leavers (43.1% male, mean age=17.14years) completed a questionnaire at Whitsunday Schoolies, Queensland, Australia on 23-24 November 2010. Nearly 49% of respondents had attended Choices. The survey included measures of alcohol use, illicit drug use and associated problems during Schoolies and a month prior to Schoolies. Results. After controlling for gender and pre-Schoolies drinking, school leavers who attended Choices were significantly less likely to report illicit drug use (OR=0.51, P<0.05) and problem behaviours (OR=0.40, P<0.01) than those who did not attend Choices. There was, however, no intervention effect in risky drinking (i.e. drank on 5 or more days, typical amount five or more standard drink and binge drank on 3 or more days) at Schoolies (OR=0.92, P=0.80). Discussion and Conclusions. Delivery of a youth-specific applied theatre prevention program employing a harm minimisation framework may be effective in reducing high-risk behaviours associated with alcohol consumption at celebratory events, even if young people expect to engage in excessive alcohol consumption. [Quek L-H, White A, Low C, Brown J, Dalton N, Dow D, Connor JP. Good choices, great future: An applied theatre prevention program to reduce alcohol-related risky behaviours during Schoolies. Drug Alcohol Rev 2012;31:897–902]

A time series analysis of presentations to Queensland health facilities for alcohol-related conditions, following the increase in 'alcopops' tax

Objective: In response to concerns about the health consequences of high-risk drinking by young people, the Australian Government increased the tax on pre-mixed alcoholic beverages ('alcopops') favoured by this demographic. We measured changes in admissions for alcohol-related harm to health throughout Queensland, before and after the tax increase in April 2008. Methods: We used data from the Queensland Trauma Register, Hospitals Admitted Patients Data Collection, and the Emergency Department Information System to calculate alcohol-related admission rates per 100,000 people, for 15 - 29 year-olds. We analysed data over 3 years (April 2006 - April 2009), using interrupted time-series analyses. This covered 2 years before, and 1 year after, the tax increase. We investigated both mental and behavioural consequences (via F10 codes), and intentional/unintentional injuries (S and T codes). Results: We fitted an auto-regressive integrated moving average (ARIMA) model, to test for any changes following the increased tax. There was no decrease in alcohol-related admissions in 15 - 29 year-olds. We found similar results for males and females, as well as definitions of alcohol-related harms that were narrow (F10 codes only) and broad (F10, S and T codes). Conclusions: The increased tax on 'alcopops' was not associated with any reduction in hospital admissions for alcohol-related harms in Queensland 15 - 29 year-olds.