Mel Ziman

Inhaled methoxyflurane and intranasal fentanyl for prehospital management of visceral pain in an Australian ambulance service

Objective This study analysed the analgesic effect and changes in vital signs associated with administration of inhaled Methoxyflurane (MTX) and/or intranasal Fentanyl (INF) for prehospital management of visceral pain. Method A retrospective, observational study reviewing 1024 randomly selected records of patients with presumed visceral pain administered MTX (465), INF (397) or both (162) by the Western Australian Ambulance Service between January 2004 and February 2006. Clinical variables assessed included systolic blood pressure, pulse rate, respiration rate and Glasgow Coma Scale score. Pain was assessed utilising Visual/Verbal Analogue Scale pain scores. Results Overall effects on vital signs appeared favourable 5 min after use and at hospital arrival with either agent alone or in combination. As sole agents, MTX produced the greatest initial pain scores reduction (2.0 (1.7 to 2.2) vs 1.6 (1.4 to 1.8)) (mean (95% CI), and INF provided greater pain reduction by hospital arrival (3.2 (2.9 to 3.5) vs 2.5 (2.1 to 2.9)). While both agents were effective, INF provided a greater pain score reduction for cardiac (3.0 (2.6 to 3.4) vs 2.3 (1.8 to 2.8)), female (3.4 (2.9 to 4.0) v 2.5 (2.0 to 3.0)) and age 75+ patients (3.2 (2.5 to 3.8) vs 1.8 (1.0 to 2.5)). Combined use of agents was not advantageous. Conclusions MTX and INF are effective agents for providing visceral pain analgesia in the prehospital setting. While MTX provided a more rapid onset of pain relief, INF provided superior analgesia after subsequent doses and in female, cardiac and older patients.

Pax3 transcripts in melanoblast development

The transcription factor encoded by PAX3 is among the first expressed in the embryo, with a key role in development of the melanocytic lineage. Re‐expression of PAX3, consistently observed in cutaneous malignant melanoma (CMM) as compared to normal melanocytes, appears linked to progression of CMM. Previous research has identified PAX3d (encoded by exons 1–9) as the predominant isoform present in CMM, together the with an alternate isoform PAX3c (encoded by exons 1–8). We investigated the expression of Pax3c and Pax3d transcripts during mouse development. The reverse transcription–polymerase chain reaction and immunohistochemistry experiments presented here implicate these transcripts in melanoblast development and demonstrate significant spatial and temporal differences in their expression. Differences in expression were also noted during active hair regrowth in adult skin, which is accompanied by proliferation and migration of melanoblasts into the hair cortex to color new hair. Results indicate that the defined spatial and temporal expression of Pax3d may be linked to either melanoblast proliferation or migration during melanogenesis.

Advances in personalized targeted treatment of metastatic melanoma and non-invasive tumor monitoring

Despite extensive scientific progress in the melanoma field, treatment of advanced stage melanoma with chemotherapeutics and biotherapeutics has rarely provided response rates higher than 20%. In the past decade, targeted inhibitors have been developed for metastatic melanoma, leading to the advent of more personalized therapies of genetically characterized tumors. Here we review current melanoma treatments and emerging targeted molecular therapies. In particular we discuss the mutant BRAF inhibitors Vemurafenib and Dabrafenib, which markedly inhibit tumor growth and advance patients’ overall survival. However this response is almost inevitably followed by complete tumor relapse due to drug resistance hampering the encouraging initial responses. Several mechanisms of resistance within and outside the MAPK pathway have now been uncovered and have paved the way for clinical trials of combination therapies to try and overcome tumor relapse. It is apparent that personalized treatment management will be required in this new era of targeted treatment. Circulating tumor cells (CTCs) provide an easily accessible means of monitoring patient relapse and several new approaches are available for the molecular characterization of CTCs. Thus CTCs provide a monitoring tool to evaluate treatment efficacy and early detection of drug resistance in real time. We detail here how advances in the molecular analysis of CTCs may provide insight into new avenues of approaching therapeutic options that would benefit personalized melanoma management.

The role of Pax7 in determining the cytoarchitecture of the superior colliculus

Pax genes are a family of transcriptional regulators vital for embryonic development. One member of the family, Pax7, functions early in neural development to establish dorsal polarity of the neural tube, and continuous refinement of its expression affords regional identity to brain nuclei, in particular the superior colliculus. Pax7 expression within the superior colliculus is eventually restricted to the stratum griseum et fibrosum superficiale (SGFS), the retinorecipient layer to which the optic nerve projects. The key role of Pax7 in specification of the superior colliculus has been highlighted by misexpression studies which result in ectopic formation of superior collicular tissue with characteristic laminae innervated by retinal ganglion cell axons. Here we review the role of Pax7 in formation of the superior colliculus and discuss the possibility that Pax7 may also assist in refinement of correct topographic mapping.

Isolation and detection of circulating tumour cells from metastatic melanoma patients using a slanted spiral microfluidic device

Circulating Tumour Cells (CTCs) are promising cancer biomarkers. Several methods have been developed to isolate CTCs from blood samples. However, the isolation of melanoma CTCs is very challenging as a result of their extraordinary heterogeneity, which has hindered their biological and clinical study. Thus, methods that isolate CTCs based on their physical properties, rather than surface marker expression, such as microfluidic devices, are greatly needed in melanoma. Here, we assessed the ability of the slanted spiral microfluidic device to isolate melanoma CTCs via label-free enrichment. We demonstrated that this device yields recovery rates of spiked melanoma cells of over 80% and 55%, after one or two rounds of enrichment, respectively. Concurrently, a two to three log reduction of white blood cells was achieved with one or two rounds of enrichment, respectively. We characterised the isolated CTCs using multimarker flow cytometry, immunocytochemistry and gene expression. The results demonstrated that CTCs from metastatic melanoma patients were highly heterogeneous and commonly expressed stem-like markers such as PAX3 and ABCB5. The implementation of the slanted microfluidic device for melanoma CTC isolation enables further understanding of the biology of melanoma metastasis for biomarker development and to inform future treatment approaches.Circulating Tumour Cells (CTCs) are promising cancer biomarkers. Several methods have been developed to isolate CTCs from blood samples. However, the isolation of melanoma CTCs is very challenging as a result of their extraordinary heterogeneity, which has hindered their biological and clinical study. Thus, methods that isolate CTCs based on their physical properties, rather than surface marker expression, such as microfluidic devices, are greatly needed in melanoma. Here, we assessed the ability of the slanted spiral microfluidic device to isolate melanoma CTCs via label-free enrichment. We demonstrated that this device yields recovery rates of spiked melanoma cells of over 80% and 55%, after one or two rounds of enrichment, respectively. Concurrently, a two to three log reduction of white blood cells was achieved with one or two rounds of enrichment, respectively. We characterised the isolated CTCs using multimarker flow cytometry, immunocytochemistry and gene expression. The results demonstrated that CTCs from metastatic melanoma patients were highly heterogeneous and commonly expressed stem-like markers such as PAX3 and ABCB5. The implementation of the slanted microfluidic device for melanoma CTC isolation enables further understanding of the biology of melanoma metastasis for biomarker development and to inform future treatment approaches.

Graft outcomes influenced by co-expression of Pax7 in graft and host tissue

Cell replacement therapies offer promise in the treatment of neurotrauma and neurodegenerative disorders and have concentrated on the use of primary fetal brain tissue. However, there is a growing promise of using neural stem cells, in which case other factors may be important in their successful engraftment. We therefore investigated whether the co‐expression of the major developmental transcription factor (Pax7 in this study) of donor tissue to graft site influences transplant survival and differentiation in the rat midbrain. Neural progenitor cells were prepared from either the Pax7‐expressing dorsal (DM) or non‐Pax7‐expressing ventral mesencephalon (VM) of embryonic EGFP+/+ rats. Cells were dissociated and grafted into the adult rat superior colliculus (SC) lesioned with quinolinic acid 3 days previously, a time shown to be associated with the up‐regulation of Pax7. Grafts were then examined 4 weeks later. Our results suggest the origin of the graft tissue did not alter graft survival in the SC; however, dorsal grafts appear to have a higher incidence of neuronal survival, whereas ventral grafts have a higher incidence of astrocytic survivors.

Students Reflecting on Test Performance and Feedback: An On-Line Approach.

Undergraduate students accessing on-line tests in Human Biology in three Western Australian universities were asked to complete an on-line post-test reflective survey about their perceptions of their test performance in light of automated feedback. The survey allowed pre-determined choices and comment text boxes relating to students’ perceptions of their performance, self-identified areas of difficulty and suggested strategies for improving test performance. One-third of students undertaking on-line tests responded to the optional survey, and 60% of respondents thought reflecting on feedback was useful. Students reflecting on formative rather than summative assessment reported a more strategic approach to testing, often using it to assess their knowledge and prepare for future assessment. Their reflections were more internally focused on motivation and preparation compared with those assessed summatively. Respondents were more likely to be female, older, more experienced learners who had scored well in the test. Younger respondents expected higher scores than they achieved and were less likely to reflect, but, when they did, were more likely to select pre-determined reasons for their performance and less likely to suggest strategies for improvement. These results support formal training and scaffolded integration of reflection into on-line assessment feedback, especially for less experienced learners.

A diagnostic autoantibody signature for primary cutaneous melanoma

Melanoma is an aggressive form of skin cancer that is curable by surgical excision in the majority of cases, if detected at an early stage. To improve early stage melanoma detection, the development of a highly sensitive diagnostic test is of utmost importance. Here we aimed to identify antibodies to a panel of tumour associated antigens that can differentiate primary melanoma patients and healthy individuals. A total of 245 sera from primary melanoma patients and healthy volunteers were screened against a high-throughput microarray platform containing 1627 functional proteins. Following rigorous statistical analysis, we identified a combination of 10 autoantibody biomarkers that, as a panel, displays a sensitivity of 79%, specificity of 84% and an AUC of 0.828 for primary melanoma detection. This melanoma autoantibody signature may prove valuable for the development of a diagnostic blood test for routine population screening that, when used in conjunction with current melanoma diagnostic techniques, could improve the early diagnosis of this malignancy and ultimately decrease the mortality rate of patients.

M17 The effect of multidisciplinary therapy on objective and subjective sleep quality in premanifest huntington’s disease

Background Subjective and objective sleep disturbances have been reported in premanifest Huntington’s disease (pre-HD). These disturbances have the potential to induce structural and functional changes to the brain, including cognitive deficits, and may facilitate, or even contribute to, disease onset and progression. Treatment of sleep disturbances could therefore improve function in affected individuals. Here, we investigated the effects of a non-pharmaceutical, short duration multidisciplinary therapy intervention on subjective and objective sleep in pre-HD participants. Aims To evaluate the effects of multidisciplinary therapy on sleep patterns and subjective sleep quality in pre-HD participants. Methods Eleven pre-HD participants aged 40 ± 11 years (5 male) underwent a twelve week multidisciplinary therapy intervention consisting of three, two hour sessions per week encompassing supervised group exercise and cognitive training. Objective (actigraphy) and subjective (Pittsburgh Sleep Quality Index, Epworth Sleepiness Scale, Consensus Sleep Diary) measures of sleep were obtained at baseline, six weeks and twelve weeks. Stress, depression and anxiety were measured at each time-point using the Perceived Stress Scale and the Hospital Anxiety and Depression Scale (as potential confounders of sleep abnormalities). Results All participants completed the required number of therapy sessions (30 sessions in total) for the study. Decreases, relative to baseline, in sleep onset latency (d > 0.5) and number and duration of awakenings (d > 0.2) were observed following the 12 week intervention. Subjective sleep quality also improved following 12 weeks of physical and cognitive training (d > 0.5). Conclusion This study provides evidence that multidisciplinary therapy has the potential to improve subjective and objective sleep quality in pre-HD. It is possible that a longer intervention duration may further improve sleep quality outcomes in this population.

M14 The effect of multidisciplinary therapy on cognition in premanifest huntington’s disease: an exploratory study

Background Despite immense scientific efforts, there are still no proven drug agents that delay or treat cognitive impairments in individuals with Huntington’s disease (HD). Recent clinical and preclinical data suggests that environmental interventions have a positive impact on many cognitive domains that are affected by HD. There are, however, no empirical studies on the effect of environmental interventions on cognition in individuals with premanifest HD. Intervening during this early stage of the disease may support and/or boost cognitive abilities in affected individuals. Aims To evaluate the effectiveness of a nine month multidisciplinary therapy intervention on cognition in individuals with premanifest HD. Methods Eighteen individuals with premanifest HD (13 women, 5 men; 43 ± 13 years) undertook a multidisciplinary therapy intervention for nine months. The intervention consisted of three times weekly supervised aerobic and resistance exercise, dual task and computerised cognitive training. Participants were assessed using validated cognitive measures before and after the intervention. Results Participants completed all mandatory training sessions throughout the study. Significant improvements in verbal learning and memory (Hopkins Verbal Learning Test), information processing speed (Symbol Digit Modalities Test), attention (Trail Making Test Part A), cognitive flexibility (Trail Making Test Part B) and problem solving (One Touch Stockings of Cambridge) were observed following the intervention. Conclusions This study provides preliminary evidence on the positive effects of multidisciplinary therapy on cognition in individuals with premanifest HD. Larger controlled trials are, however, required to confirm these promising findings.