Studies of assisted reproduction in the spotted grass frog Limnodynastes tasmaniensis: ovulation, early development and microinjection (ICSI)

Assisted Reproductive Technologies (ART) offer a wide range of techniques that have the potential to augment efforts to conserve and manage endangered amphibians and improve wild and captive population numbers. Gametes and tissues of species nearing endangered or extinct status can be cryopreserved and stored in gene banks, to provide material that can be utilised in the future as ART methods are refined. The Spotted Grass Frog, Limnodynastes tasmaniensis, is an abundant amphibian species in South-Eastern Australia of the family Myobatrachidae, that is suitable for the development of ART systems that can be applied to the threatened and endangered myobatrachid and other amphibian species native to Australia. The aim of this study was to advance the understanding of ovulation, fertilisation and embryo nic development of Lim. tasmaniensis and in vitro manipulations of reproduction and development for use in the development of advanced ART procedures such as intracytoplasmic spermatozoon injection (ICSI), androgenesis and nuclear transfer. Ovulation in amphibians can be induced by protocols utilising natural or synthetic hormones. All protocols tested on Lim. tasmaniensis in this study required two injections and the most effective protocols continued to require a first injection of pituitary extracts to induce ovulation. The second injection was, however, successfully replaced by synthetic chorionic gonadotrophin at a threshold dosage of 100 iu and halved the number of cane toads required to source the pituitaries. A combination of LHRH and Pimozide offered a less effective protocol, that did not require the use of pituitary extracts, and avoided the risk of pathogen transfer associated with unsterilised pituitary extracts. Unfertilised eggs of Lim. tasmaniensis were exposed to media of various osmolalities to determine media effects on eggs and their surrounding jelly layers that might impact on egg viability and fertilisability. Osmolality had no effect upon the egg diameter, however, rapid swelling of the jelly layers occurred within 15 minutes of exposure to various media treatments and plateaued from 30-90 minutes without further expansion. Swelling of the jelly layers was increased in hypotonic media (2.5% SAR, H2O) and minimised in the isotonic media (100% SAR). The optimal conditions for the culture of Lim. tasmaniensis eggs were identified as a holding media of 100% SAR, followed by a medium change to 2.5% SAR at insemination. This sequence of media minimised the rate of swelling of the jelly layers prior to contact with the spermatozoa, and maximised the activation of spermatozoa and eggs throughout fertilisation and embryonic development. Embryos of Lim. tasmaniensis were cultured at four temperatures (13 C, 17 C, 23 C and 29 C), to determine the effect of temperature on cleavage and embryonic development rates. Embryonic development progressed through a sequence of stages that were not altered by changes in temperature. However cleavage rates were affected by changes in temperature as compared with normal embryonic growth at 23 C. Embryonic development was suspended at the lowest temperature (13 C) while embryonic viability was maintained. A moderate decrease in temperature (17 C) slowed cleavage, while the highest temperature (29 C) increased the cleavage rate, but decreased the embryo survival. Rates of embryonic development can be manipulated by changes in temperature and this method can be used to source blastomeres of a specific size/stage at a predetermined age or halt cleavage at specific stages for embryos or embryo derived cells to be included in ART procedures. This study produced the first report of the application of Intracytoplasmic Spermatozoon Injection (ICSI) in an Australian amphibian. Eggs that were activated by microinjection with a single spermatozoon (n=50) formed more deep, but abnormal, cleavage furrows post-injection (18/50, 36%), than surface changes (12/50, 24%). This result is in contrast to eggs injected without a spermatozoon (n=42), where the majority of eggs displayed limited surface changes (36/42, 86%), and few deep, abnormal furrows (3/42, 7%). Three advanced embryos (3/50, 6%) were produced by ICSI that developed to various stages within the culture system. Technical difficulties were encountered that prevented the generation of any metamorphs from ICSI tadpoles. Nevertheless, when these blocks to ICSI are overcome, the ICSI procedure will be both directly useful as an ART procedure in its own right, and the associated refinement of micromanipulation procedures will assist in the development of other ART procedures in Lim. tasmaniensis. A greater understanding of basic reproductive and developmental biology in Lim. tasmaniensis would greatly facilitate refinement of fertilisation by ICSI. Assisted Reproductive Technologies, in conjunction with gene banks may in the future regenerate extinct amphibian species, and assist in the recovery of declining amphibian populations nationally and worldwide.

A pragmatic approach to intellectual property and development : a case study of the Jordanian copyright law in the internet age

On October 4, 2004, Brazil and Argentina requested that WIPO adopt a development-oriented approach to IP and to reconsider its work in relation to developing countries. In October, 2007, WIPO member States adopted a historic decision for the benefit of developing countries, to establish a WIPO Development Agenda. Although there have been several studies related to IP and development that call for IP laws in developing countries to be development-friendly, there is little research that attempts to provide developing countries with practical measures to achieve that goal. This article takes the copyright law in Jordan as a case study and shows how, in practical terms, a pro-development-oriented approach could be implemented in the copyright laws of developing countries. It provides specific recommendations for developing countries to ensure that their IP laws are aligned with and serve their social and economic development objectives.

The platform of spoof videos : The case of Tudou.com

This paper analyses the enabling and constraining role of platforms in the development of online video-spoofing culture in China. Using Tudou.com, one of China’s most popular video-sharing sites, as a case study, the paper discusses how video-sharing sites facilitate and curtail the self, and how they negotiate and balance among users, clients and the state, teasing out the complex politics of platforms in China. Particular reference is made to three “star users”, Jiaoshou, Hu Ge and Corndog, who gained fame for themselves via the popularity of their spoof videos on this platform.

Development of a food frequency questionnaire for Sri Lankan adults

Abstract (provisional) Background Food Frequency Questionnaires (FFQs) are commonly used in epidemiologic studies to assess long-term nutritional exposure. Because of wide variations in dietary habits in different countries, a FFQ must be developed to suit the specific population. Sri Lanka is undergoing nutritional transition and diet-related chronic diseases are emerging as an important health problem. Currently, no FFQ has been developed for Sri Lankan adults. In this study, we developed a FFQ to assess the regular dietary intake of Sri Lankan adults. Methods A nationally representative sample of 600 adults was selected by a multi-stage random cluster sampling technique and dietary intake was assessed by random 24-h dietary recall. Nutrient analysis of the FFQ required the selection of foods, development of recipes and application of these to cooked foods to develop a nutrient database. We constructed a comprehensive food list with the units of measurement. A stepwise regression method was used to identify foods contributing to a cumulative 90% of variance to total energy and macronutrients. In addition, a series of photographs were included. Results We obtained dietary data from 482 participants and 312 different food items were recorded. Nutritionists grouped similar food items which resulted in a total of 178 items. After performing step-wise multiple regression, 93 foods explained 90% of the variance for total energy intake, carbohydrates, protein, total fat and dietary fibre. Finally, 90 food items and 12 photographs were selected. Conclusion We developed a FFQ and the related nutrient composition database for Sri Lankan adults. Culturally specific dietary tools are central to capturing the role of diet in risk for chronic disease in Sri Lanka. The next step will involve the verification of FFQ reproducibility and validity.

How males and females define speeding and how they’d feel getting caught for it : some implications for anti-speeding message development

Speeding represents a major contributor to road trauma, increasing crash frequency and severity. Antispeeding campaigns represent a key strategy aimed at discouraging individuals from speeding. This paper investigated salient beliefs underpinning male and female drivers’ travel speed behaviour, with the view to use such insight to, ultimately, inform the content of targeted anti-speeding messages. A survey of N = 751 (579 males, 16-79 years) drivers assessed what they regarded as speeding in 60km/hr and 100km/hr zones and their beliefs about how they would respond to receiving a speeding infringement. Participants responded to scales which extended up to 20km/hr above each respective speed limit, the lowest speed that they considered was speeding and the speed at which they would be willing to drive and still feel in control. For analyses, to enable greater scrutiny of potential gender differences regarding the speeds identified, participants’ responses to these items were categorised into 5km/hr increments and chi-square analyses conducted. For their responses to (beliefs about) the possibility of being caught speeding, drivers were asked how applicable various beliefs were to them (e.g., feeling unlucky). These beliefs were analysed via MANOVA. The results revealed that there was considerable variability in the speeds identified, thus supporting the value of categorising speeds. Within the 100km/hr zone, based on the categories, a significant difference was found regarding the speed that males would be willing to drive (and still feel in control) relative to females. Specifically, the greatest proportion of males (30.4%) identified speeds within the 106-110km/hr category whereas the greatest proportion of females (38.1%) identified a lower speed, within the 101-105km/hr category, as the speed they would be willing to drive. No other significant differences emerged, however, either in relation to the definition of speeding reported for 100km/hr zones (i.e., males and females tended to identify a similar speed as indicative of speeding) nor for these same items as assessed in relation to the 60km/hr zones. For their responses to the possibility of being caught, males were significantly more likely than females to report that, if caught, a likely response they would have would be to think that they had still been driving safely. In contrast, females were significantly more likely than males to report thinking that their speeding had been unsafe and that they should not have been speeding. Females were also significantly more likely to report feeling embarrassed to tell important others about having received a speeding infringement than males. The findings are discussed in terms of their implications for developing well-targeted advertising messages aimed at discouraging drivers’ from speeding.

Supporting the learning of rapid application development in a database environment

Intelligent Tutoring Systems (ITSs) are computer systems designed to provide individualised help to students, learning in a problem solving context. The difference between an ITS and a Computer Assisted Instruction (CAI) system is that an ITS has a Student Model which allows it to provide a better educational environment. The Student Model contains information on what the student knows, and does not know, about the domain being learnt, as well as other personal characteristics such as preferred learning style. This research has resulted in the design and development of a new ITS: Personal Access Tutor (PAT). PAT is an ITS that helps students to learn Rapid Application Development in a database environment. More specifically, PAT focuses on helping students to learn how to create forms and reports in Microsoft Access. To provide an augmented learning environment, PAT’s architecture is different to most other ITSs. Instead of having a simulation, PAT uses a widelyused database development environment (Microsoft Access). This enables the students to ask for help, while developing real applications using real database software. As part of this research, I designed and created the knowledge base required for PAT. This contains four models: the domain, student, tutoring and exercises models. The Instructional Expert I created for PAT provides individualised help to the students to help them correctly finish each exercise, and also proposes the next exercise that a student should work on. PAT was evaluated by students enrolled in the Databases subject at QUT, and by staff members involved in teaching the subject. The results of the evaluation were positive and are discussed in the thesis.

The development of a supportive care needs assessment tool for Indigenous people with cancer

Background: Little is known about the supportive care needs of Indigenous people with cancer and to date, existing needs assessment tools have not considered cultural issues for this population. We aimed to adapt an existing supportive care needs assessment tool for use with Indigenous Australians with cancer. Methods: Face-to-face interviews with Indigenous cancer patients (n = 29) and five focus groups with Indigenous key-informants (n = 23) were conducted to assess the face and content validity, cultural acceptability, utility and relevance of the Supportive Care Needs Survey - Short Form 34 (SCNS-SF34) for use with Indigenous patients with cancer. Results: All items from the SCNS-SF34 were shortened and changed to use more appropriate language (e.g. the word 'anxiety' was substituted with 'worry'). Seven questions were omitted (e.g. items on death and future considerations) as they were deemed culturally inappropriate or irrelevant and 12 items were added (e.g. accessible transport). Optional instructions were added before the sexual items. The design and response format of the SCNS-SF34 was modified to make it easier to use for Indigenous cancer patients. Given the extensive modifications to the SCNS-SF34 and the liklihood of a different factor structure we consider this tool to be a new tool rather than a modification. The Supportive care needs assessment tool for Indigenous people (SCNAT-IP) shows promising face and content validity and will be useful in informing services where they need to direct their attention for these patients. Conclusions: Indigenous people with cancer have language, customs and specific needs that are not accommodated within the standard SCNS-SF34. Our SCNAT-IP improves acceptability, relevance and face validity for Indigenous-specific concerns. Our SCNAT-IP will allow screening for supportive care needs that are specific to Indigenous cancer patients' and greatly inform targeted policy development and practice.

Travel context : development of a model to measure destination brand performance across different travel situations

Purpose: There is a lack of theory relating to destination brand performance measurement in the destination branding literature, which emerged in the late 1990s (see for example Dosen, Vransevic, & Prebezac, 1998). Additionally, there is a lack of research about the importance of travel context in consumers’ destination decision making (Hu & Ritchie, 1993). This study develops a structural model to measure destination brand performance across different travel situations. The theory of planned behaviour (TpB) was utilised as a framework to underpin the consumer-based brand equity (CBBE) hierarchy to develop a model of destination brand performance. Research approach: A proposed model of destination brand performance was developed through a review of the literature. The first study was used to identify destination image attributes (the core construct) using an analysis of the literature, a document analysis, and personal interviews using the Repertory Test qualitative technique. Underpinned by Personal Construct Theory (PCT), the Repertory Test enables the elicitation of attributes consumers use to evaluate destinations when considering travel. Data was examined in the first study to i) identify any attribute differences in travel contexts and ii) create a scale for use in a questionnaire. A second study was conducted to test the proposed model using a questionnaire with eight groups of participants to assess four destinations across two travel contexts. The model was tested utilising structural equation modelling. Findings: The first study resulted in a list of 29 destination image attributes for use in a scale index. Attributes were assessed across travel contexts and few differences were identified. The second study assessed the congruence of destination brand identity (the destination marketing organisation’s desired image) and destination brand image (the actual perceptions held by consumers) using importance-performance analyses. Finally, the proposed model of destination brand performance was tested. Overall the data supported the model of destination brand performance across travel contexts and destinations. Additionally, this was compared to consumers’ decision sets, further supporting the model. Value: This research provides a contribution to the destination marketing literature through the development of a measurement of destination brand performance underpinned by TpB. Practically; it will provide destination marketing organisations with a tool to track destination brand performance, relative to key competing places, over time. This is important given the development of a destination brand is a long term endeavour.

Rate of torque and electromyographic development during anticipated eccentric contraction is lower in previously strained hamstrings

Background: Hamstring strain injuries are prevalent in sport and re-injury rates have been high for many years. Whilst much focus has centred on the impact of previous hamstring strain injury on maximal eccentric strength, high rates of torque development is also of interest, given the important role of the hamstrings during the terminal swing phase of running. The impact of prior strain injury on myoelectrical activity of the hamstrings during tasks requiring high rates of torque development has received little attention. Purpose: To determine if recreational athletes with a history of unilateral hamstring strain injury, who have returned to training and competition, will exhibit lower levels of myoelectrical activity during eccentric contraction, rate of torque development and impulse 30, 50 and 100ms after the onset of myoelectrical activity or torque development in the previously injured limb compared to the uninjured limb. Study design: Case-control study Methods: Twenty-six recreational athletes were recruited. Of these, 13 athletes had a history of unilateral hamstring strain injury (all confined to biceps femoris long head) and 13 had no history of hamstring strain injury. Following familiarisation, all athletes undertook isokinetic dynamometry testing and surface electromyography assessment of the biceps femoris long head and medial hamstrings during eccentric contractions at -60 and -1800.s-1. Results: In the injured limb of the injured group, compared to the contralateral uninjured limb rate of torque development and impulse was lower during -600.s-1 eccentric contractions at 50 (RTD, injured limb = 312.27 ± 191.78Nm.s-1 vs. uninjured limb = 518.54 ± 172.81Nm.s-1, p=0.008; IMP, injured limb = 0.73 ± 0.30 Nm.s vs. uninjured limb = 0.97 ± 0.23 Nm.s, p=0.005) and 100ms (RTD, injured limb = 280.03 ± 131.42Nm.s-1 vs. uninjured limb = 460.54.54 ± 152.94Nm.s-1,p=0.001; IMP, injured limb = 2.15 ± 0.89 Nm.s vs. uninjured limb = 3.07 ± 0.63 Nm.s, p<0.001) after the onset of contraction. Biceps femoris long head muscle activation was lower at 100ms at both contraction speeds (-600.s-1, normalised iEMG activity (x1000), injured limb = 26.25 ± 10.11 vs. uninjured limb 33.57 ± 8.29, p=0.009; -1800.s-1, normalised iEMG activity (x1000), injured limb = 31.16 ± 10.01 vs. uninjured limb 39.64 ± 8.36, p=0.009). Medial hamstring activation did not differ between limbs in the injured group. Comparisons in the uninjured group showed no significant between limbs difference for any variables. Conclusion: Previously injured hamstrings displayed lower rate of torque development and impulse during slow maximal eccentric contraction compared to the contralateral uninjured limb. Lower myoelectrical activity was confined to the biceps femoris long head. Regardless of whether these deficits are the cause of or the result of injury, these findings could have important implications for hamstring strain injury and re-injury. Particularly, given the importance of high levels of muscle activity to bring about specific muscular adaptations, lower levels of myoelectrical activity may limit the adaptive response to rehabilitation interventions and suggest greater attention be given to neural function of the knee flexors following hamstring strain injury.

Impact of previous hamstring strain injury on rate of torque and EMG development during eccentric contraction.

Background: Hamstring strain injuries (HSIs) are prevalent in sport and re-injury rates have been high for many years. Whilst much focus has centred on the impact of previous hamstring strain injury on maximal eccentric strength, high rates of torque development is also of interest, given the important role of the hamstrings during the terminal swing phase of gait. The impact of prior strain injury on neuromuscular function of the hamstrings during tasks requiring high rates of torque development has received little attention. The purpose of this study is to determine if recreational athletes with a history of unilateral hamstring strain injury, who have returned to training and competition, will exhibit lower levels of eccentric muscle activation, rate of torque development and impulse 30, 50 and 100ms after the onset of electromyographical or torque development in the previously injured limb compared to the uninjured limb. Methods: Twenty-six recreational athletes were recruited. Of these, 13 athletes had a history of unilateral hamstring strain injury (all confined to biceps femoris long head) and 13 had no history of hamstring strain injury. Following familiarisation, all athletes undertook isokinetic dynamometry testing and surface electromyography assessment of the biceps femoris long head and medial hamstrings during eccentric contractions at -60 and -1800.s-1. Results: In the injured limb of the injured group, compared to the contralateral uninjured limb rate of torque development and impulse was lower during -600.s-1 eccentric contractions at 50 (RTD, p=0.008; IMP, p=0.005) and 100ms (RTD, p=0.001; IMP p<0.001) after the onset of contraction. There was also a non-significant trend for rate of torque development during -1800.s-1 to be lower 100ms after onset of contraction (p=0.064). Biceps femoris long head muscle activation was lower at 100ms at both contraction speeds (-600.s-1, p=0.009; -1800.s-1, p=0.009). Medial hamstring activation did not differ between limbs in the injured group. Comparisons in the uninjured group showed no significant between limbs difference for any variables. Conclusion: Previously injured hamstrings displayed lower rate of torque development and impulse during eccentric contraction. Lower muscle activation was confined to the biceps femoris long head. Regardless of whether these deficits are the cause of or the result of injury, these findings have important implications for hamstring strain injury and re-injury and suggest greater attention be given to neural function of the knee flexors.

Intellectual property and development : theory and practice

Since the 1960s, many developing countries have introduced IP laws to help them in their social and economic development. Introducing these laws was considered as a civilised act and a precondition of developing countries‘ progress from being =under-developed‘ to becoming =developed‘. In 2004, Brazil and Argentina presented a comprehensive proposal on behalf of developing countries to establish the Development Agenda in the World Intellectual Property Organisation (WIPO). They put forward a view that IP laws in their current form are not helping those countries in their development, as is constantly being suggested by developed countries, and that there is a need to rethink the international IP system and the work of WIPO. The research undertaken examines the correlation between IP and social and economic development. It investigates how IP systems in developing countries could work to advance their development, especially in the context of the internet. The research considers the theory and practice of IP and development, and proposes a new IP framework which developing countries could employ to further their social and economic development.

Refugees : the Millenium Development Goals' overlooked priority group

The Millennium Development Goals (MDGs) apply to the world’s 43 million refugees and forcibly displaced. While States have an obligation to meet minimum human rights levels for all persons including 'non-citizens', UNHCR must ensure countries adopting MDG targets report on the progress of their refugee populations. If we are to make significant change within the MDG time-frame, the health and human rights needs of refugees and the displaced must be urgently integrated into the development policy agendas of sovereign States, and be at the fore of the international community’s attention.

Plant made anti-HIV microbicides-A field of opportunity

HIV remains a significant global burden and without an effective vaccine, it is crucial to develop microbicides to halt the initial transmission of the virus. Several microbicides have been researched with various levels of success. Amongst these, the broadly neutralising antibodies and peptide lectins are promising in that they can immediately act on the virus and have proven efficacious in in vitro and in vivo protection studies. For the purpose of development and access by the relevant population groups, it is crucial that these microbicides be produced at low cost. For the promising protein and peptide candidate molecules, it appears that current production systems are overburdened and expensive to establish and maintain. With recent developments in vector systems for protein expression coupled with downstream protein purification technologies, plants are rapidly gaining credibility as alternative production systems. Here we evaluate the advances made in host and vector system development for plant expression as well as the progress made in expressing HIV neutralising antibodies and peptide lectins using plant-based platforms. © 2012 Elsevier Inc.

Replicative intermediates of maize streak virus found during leaf development

Geminiviruses of the genera Begomovirus and Curtovirus utilize three replication modes: complementary-strand replication (CSR), rolling-circle replication (RCR) and recombinationdependent replication (RDR). Using two-dimensional gel electrophoresis, we now show for the first time that maize streak virus (MSV), the type member of the most divergent geminivirus genus, Mastrevirus, does the same. Although mastreviruses have fewer regulatory genes than other geminiviruses and uniquely express their replication-associated protein (Rep) from a spliced transcript, the replicative intermediates of CSR, RCR and RDR could be detected unequivocally within infected maize tissues. All replicative intermediates accumulated early and, to varying degrees, were already present in the shoot apex and leaves at different maturation stages. Relative to other replicative intermediates, those associated with RCR increased in prevalence during leaf maturation. Interestingly, in addition to RCR-associated DNA forms seen in other geminiviruses, MSV also apparently uses dimeric open circular DNA as a template for RCR. © 2010 SGM.