Development of novel mass spectrometry methodology to detect post-translational modifications in oxidative stress and disease

This paper presented at the European Meeting of the Society-for-Free-Radical-Research-Europe 2007, discusses the development of novel mass spectrometry methodology to detect post-translational modifications in oxidative stress and disease.

Post-Traumatic Stress Disorder (PTSD) and Its relationship with perinatal bereavement: definitions, reactions, adjustments, and grief.

A small percentage of women present with diagnosed PTSD post childbirth; however, though this an area of increasing research and clinical interest generally, little is known about PTSD in relation to perinatal bereavement. Health professionals must however be aware of the mechanisms and models of loss following perinatal bereavement in order to identify potential mental health phenomena which may be implicated in the development of PTSD symptomology. Understanding the predictive factors which may give early warning signs is an important component of the clinical evidence base. This chapter discusses mechanisms, models, and risk factors in relation to perinatal bereavement and the development of PTSD.

The manipulation of apoptosis-related genes to generate resistance to Fusarium wilt and water stress in banana

Bananas are susceptible to a diverse range of biotic and abiotic stresses, many of which cause serious production constraints worldwide. One of the most destructive banana diseases is Fusarium wilt caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense (Foc). No effective control strategy currently exists for this disease which threatens global banana production. Although disease resistance exists in some wild bananas, attempts to introduce resistance into commercially acceptable bananas by conventional breeding have been hampered by low fertility, long generation times and association of poor agronomical traits with resistance genes. With the advent of reliable banana transformation protocols, molecular breeding is now regarded as a viable alternative strategy to generate disease-resistant banana plants. Recently, a novel strategy involving the expression of anti-apoptosis genes in plants was shown to result in resistance against several necrotrophic fungi. Further, the transgenic plants showed increased resistance to a range of abiotic stresses. In this thesis, the use of anti-apoptosis genes to generate transgenic banana plants with resistance to Fusarium wilt was investigated. Since water stress is an important abiotic constraint to banana production, the resistance of the transgenic plants to water stress was also examined. Embryogenic cell suspensions (ECS) of two commercially important banana cultivars, Grand Naine (GN) and Lady Finger (LF), were transformed using Agrobacterium with the anti-apoptosis genes, Bcl-xL, Bcl-xL G138A, Ced-9 and Bcl- 2 3’ UTR. An interesting, and potentially important, outcome was that the use of anti-apoptosis genes resulted in up to a 50-fold increase in Agrobacterium-mediated transformation efficiency of both LF and GN cells over vector controls. Regenerated plants were subjected to a complete molecular characterisation in order to detect the presence of the transgene (PCR), transcript (RT-PCR) and gene product (Western blot) and to determine the gene copy number (Southern blot). A total of 36 independently-transformed GN lines (8 x Bcl-xL, 5 x Bcl-xL G138A, 15 x Ced-9 and 8 x Bcl-2 3’ UTR) and 41 independently-transformed LF lines (8 x Bcl-xL, 7 x BclxL G138A, 13 x Ced-9 and 13 x Bcl-2 3’ UTR) were identified. The 41 transgenic LF lines were multiplied and clones from each line were acclimatised and grown under glasshouse conditions for 8 weeks to allow monitoring for phenotypic abnormalities. Plants derived from 3 x Bcl-xL, 2 x Ced-9 and 5 x Bcl-2 3’ UTR lines displayed a variety of aberrant phenotypes. However, all but one of these abnormalities were off-types commonly observed in tissue-cultured, non-transgenic banana plants and were therefore unlikely to be transgene-related. Prior to determining the resistance of the transgenic plants to Foc race 1, the apoptotic effects of the fungus on both wild-type and Bcl-2 3’ UTR-transgenic LF banana cells were investigated using rapid in vitro root assays. The results from these assays showed that apoptotic-like cell death was elicited in wild-type banana root cells as early as 6 hours post-exposure to fungal spores. In contrast, these effects were attenuated in the root cells of Bcl-2 3’ UTR-transgenic lines that were exposed to fungal spores. Thirty eight of the 41 transgenic LF lines were subsequently assessed for resistance to Foc race 1 in small-plant glasshouse bioassays. To overcome inconsistencies in rating the internal (vascular discolouration) disease symptoms, a MatLab-based computer program was developed to accurately and reliably assess the level of vascular discolouration in banana corms. Of the transgenic LF banana lines challenged with Foc race 1, 2 x Bcl-xL, 3 x Ced-9, 2 x Bcl-2 3’ UTR and 1 x Bcl-xL G138A-transgenic line were found to show significantly less external and internal symptoms than wild-type LF banana plants used as susceptible controls at 12 weeks post-inoculation. Of these lines, Bcl-2 3’ UTR-transgenic line #6 appeared most resistant, displaying very mild symptoms similar to the wild-type Cavendish banana plants that were included as resistant controls. This line remained resistant for up to 23 weeks post-inoculation. Since anti-apoptosis genes have been shown to confer resistance to various abiotic stresses in other crops, the ability of these genes to confer resistance against water stress in banana was also investigated. Clonal plants derived from each of the 38 transgenic LF banana plants were subjected to water stress for a total of 32 days. Several different lines of transgenic plants transformed with either Bcl-xL, Bcl-xL G138A, Ced-9 or Bcl-2 3’ UTR showed a delay in visual water stress symptoms compared with the wild-type control plants. These plants all began producing new growth from the pseudostem following daily rewatering for one month. In an attempt to determine whether the protective effect of anti-apoptosis genes in transgenic banana plants was linked with reactive oxygen species (ROS)-associated programmed cell death (PCD), the effect of the chloroplast-targeting, ROS-inducing herbicide, Paraquat, on wild-type and transgenic LF was investigated. When leaf discs from wild-type LF banana plants were exposed to 10 ìM Paraquat, complete decolourisation occurred after 48 hours which was confirmed to be associated with cell death and ROS production by trypan blue and 3,3-diaminobenzidine (DAB) staining, respectively. When leaf discs from the transgenic lines were exposed to Paraquat, those derived from some lines showed a delay in decolourisation, suggesting only a weak protective effect from the transgenes. Finally, the protective effect of anti-apoptosis genes against juglone, a ROS-inducing phytotoxin produced by the causal agent of black Sigatoka, Mycosphaerella fijiensis, was investigated. When leaf discs from wild-type LF banana plants were exposed to 25 ppm juglone, complete decolourisation occurred after 48 hours which was again confirmed to be associated with cell death and ROS production by trypan blue and DAB staining, respectively. Further, TdT-mediated dUTP nick-end labelling (TUNEL) assays on these discs suggested that the cell death was apoptotic. When leaf discs from the transgenic lines were exposed to juglone, discs from some lines showed a clear delay in decolourisation, suggesting a protective effect. Whether these plants are resistant to black Sigatoka is unknown and will require future glasshouse and field trials. The work presented in this thesis provides the first report of the use of anti-apoptosis genes as a strategy to confer resistance to Fusarium wilt and water stress in a nongraminaceous monocot, banana. Such a strategy may be exploited to generate resistance to necrotrophic pathogens and abiotic stresses in other economically important crop plants.

Reducing the risk of posttraumatic stress disorder in children following natural disasters

A significant number of children suffer long term psychological disturbance following exposure to a natural disaster. Evidence suggests that a dose-response relationship exists, so that children and adolescents who experience the most intense or extensive exposure to the risk factors for PTSD are likely to develop the most serious and persistent symptoms. Risk factors include gender, age, personality, extent of exposure to the natural disaster, amount of damage to property and infrastructure, witnessing injury or death of others or perceiving a threat to their own life. Knowing these factors enables various strategies to be put in place to decrease the risk of psychological disturbance following the aftermath of traumatic events. Re-establishing a sense of safety, security and normality is important in the aftermath of a natural disaster, and promoting social connectedness, positive family functioning, and effective coping mechanisms can make children more resilient in the face of catastrophic events. This paper examines the risk and protective factors associated with the development of post traumatic stress disorder (PTSD), and considers how schools can use this knowledge to contribute to the recovery effort, and reduce the prevalence of PTSD amongst pupils in the wake of a natural disaster.

Sudanese refugees in Australia the impact of acculturation stress

Refugees from Sudan are the fastest growing community in Australia. Australian mental health professionals have to be prepared to offer services to this ethnic group along with the other mainstream and diverse consumers. In order to offer culturally competent services, these mental health professionals are required to be familiar with this emerging community. As such a review was undertaken with two main goals. Firstly, the review aimed to educate Australian mental health professionals about the demographics and culture of Sudan, the traumas encountered as a result of the civil war, factors leading to massive exodus, and the difficulties of the transit and post migration phase. Secondly, the review intended to inform Australian mental health professionals about the possible acculturation stress that is manifested in the form of intergeneration and role conflict and marital difficulties. The review highlights that there are few studies addressing acculturation stress of Sudanese refugees, and even fewer on the impact it has on relationships. Future research directions are discussed.

An examination of the base rates of post-concussion symptoms : the influence of demographics and depression

Post-concussion syndrome (PCS) is a controversial constellation of cognitive, emotional, and physical symptoms that some patients experience following a mild traumatic brain injury or concussion. PCS-like symptoms are commonly found in individuals with depression, pain, and stress, as well as healthy individuals. This study investigated the base rate of PCS symptoms in a healthy sample of 96 participants and examined the relationship between these symptoms, depression, and sample demographics. PCS symptoms were assessed using the British-Columbia Post-Concussion Symptom Inventory. Depression was measured using the Beck Depression Inventory II. Results demonstrated that: The base rate of PCS was very high; there was a strong positive relationship between depression and PCS; and demographic characteristics were not related to PCS in this sample. These findings are broadly consistent with literature suggesting a significant role for non-neurological factors in the expression of PCS symptomatology. This study adds to the growing body of literature that calls for caution in the clinical interpretation of results from PCS symptom inventories.

Mental health of newly arrived Burmese refugees in Australia : contributions of pre-migration and post-migration experience

Objective: This study documents the mental health status of people from Burmese refugee backgrounds, recently arrived in Australia; then examines the contributions of gender, premigration and postmigration factors in predicting mental health. Method: Structured interviews, including a demographic questionnaire, the Harvard Trauma Questionnaire, Postmigration Living Difficulties Checklist and Hopkins Symptom Checklist assessed premigration trauma, postmigration living difficulties, depression, anxiety, somatisation and traumatisation symptoms in a sample of 70 adults across five Burmese ethnic groups. Results: Substantial proportions of participants reported psychological distress in symptomatic ranges including: posttraumatic stress disorder (9%); anxiety (20%), and; depression (36%), as well as significant symptoms of somatisation (37%). Participants reported multiple and severe premigration traumas. Postmigration living difficulties of greatest concern included communication problems and worry about family not in Australia. Gender did not predict mental health. Level of exposure to traumatic events and postmigration living difficulties each made unique and relatively equal contributions to traumatisation symptoms. Postmigration living difficulties made unique contributions to depression, anxiety and somatisation symptoms. Conclusions: While exposure to traumatic events impacted on participants’ mental wellbeing, postmigration living difficulties had greater salience in predicting mental health outcomes of people from Burmese refugee backgrounds. Reported rates of posttraumatic stress disorder symptoms were consistent with a large review of adults across seven western countries. High levels of somatisation pointed to a nuanced expression of distress. Findings have implications for service provision in terms of implementing appropriate interventions to effectively meet the needs of this newly arrived group in Australia.

Acculturation stress in South Sudanese refugees : impact on marital relationships

A qualitative approach was used to explore the impact of acculturation stress on the marital relationships of South Sudanese refugees settled in Brisbane, Australia. Thirteen refugees, who were currently or previously married, participated in three gender specific focus groups. The perceived causes and possible solutions of conflict were thoroughly explored. Hypothetical scenarios were used to facilitate group discussion. Major issues causing conflict between couples were identified as: the management of finances and lack of family and social support. Several other areas of acculturation stress also emerged as factors associated with marital stress. There was a dissonance regarding the adherence to cultural gender roles. Freedom provided to women in Australia caused tension between the couples. Law enforcement officers were perceived as lacking cultural understanding and misinterpreting the couple distress. Finally, limited information provided to refugees pre and post migration was considered to hinder adjustment. The participants suggested a number of practical solutions to these issues which are potentially useful in guiding future refugee settlement programs.

The effect of post-match alcohol ingestion on recovery from competitive Rugby League matches

This study investigated the effects of alcohol ingestion on lower body strength and power, and physiological and cognitive recovery following competitive Rugby League matches. Nine male Rugby players participated in two matches, followed by one of two randomized interventions; a control or alcohol ingestion session. Four hours post-match, participants consumed either beverages containing a total of 1g of ethanol per kg bodyweight (vodka and orange juice; ALC) or a caloric and taste matched non-alcoholic beverage (orange juice; CONT). Pre, post, 2 h post and 16 h post match measures of countermovement jump (CMJ), maximal voluntary contraction(MVC), voluntary activation (VA), damage and stress markers of creatine kinase (CK), C-reactive protein (CRP), cortisol, and testosterone analysed from venous blood collection, and cognitive function (modified Stroop test) were determined. Alcohol resulted in large effects for decreased CMJ height(-2.35 ± 8.14 and -10.53 ± 8.36 % decrement for CONT and ALC respectively; P=0.15, d=1.40), without changes in MVC (P=0.52, d=0.70) or VA (P=0.15, d=0.69). Furthermore, alcohol resulted in a significant slowing of total time in a cognitive test (P=0.04, d=1.59), whilst exhibiting large effects for detriments in congruent reaction time (P=0.19, d=1.73). Despite large effects for increased cortisol following alcohol ingestion during recovery (P=0.28, d=1.44), post-match alcohol consumption did not unduly affect testosterone (P-0.96, d=0.10), CK (P=0.66, d=0.70) or CRP(P=0.75, d=0.60). It appears alcohol consumption during the evening following competitive rugby matches may have some detrimental effects on peak power and cognitive recovery the morning following a Rugby League match. Accordingly, practitioners should be aware of the potential associated detrimental effects of alcohol consumption on recovery and provide alcohol awareness to athletes at post-match functions.