Characterisation of Burn Blister Fluid Using Quantitative Proteomics to Aid Clinical Diagnosis

Mr Tuo Zang1,2,3, Dr Leila Cuttle1,2,4, Dr Daniel Broszczak1,2,5, Dr James Broadbent1,2,6, Ms Catherine  Tanzer1,2,4, Dr Tony Parker1,2

1Institute of Health and Biomedical Innovation, Kelvin Grove, Australia, 2Queensland University Of Technology, Kelvin Grove, Australia, 3Wound Management Innovation Co-operative Research Centre, Brisbane, Australia, 4Institute of Health and Biomedical Innovation at the Centre for Children’s Health Research, South Brisbane, Australia, 5Australian Catholic University, Banyo, Australia, 6Sciex P/L, Brisbane, Australia

Abstract:

Blister fluid (BF) is a sample matrix that can reflect both systemic and local microenvironment responses to burn injury. Our aim was to investigate the proteome of burn blister fluid using liquid chromatography tandem mass spectrometry (LC-MS/MS), which provides an objective and quantitative measurement of biomolecules in the wound site. This enables a non-invasive investigation of the cutaneous biological response to different burn severities, which may aid in the clinical diagnosis and monitor the response of treatment.

In order to generate a comprehensive peptide spectral library for quantitative information acquisition, a subset of BF samples were pooled according to burn depth (12 superficial, 12 deep-partial thickness, and 4 full thickness) and fractionated by four different methods, prior to LC-MS/MS analysis. Subsequently, all individual BF samples (n=87) were analysed using LC-MS/MS in quantitative mode (SWATH) to obtain the protein abundance. More than 800 individual proteins were identified and formed the basis of a BF protein library. Finally, more than 600 proteins in each individual sample was quantitatively profiled. This proteomic data was correlated with different clinical parameters, such as burn depth, time to re-epithelialisation, first aid, and burn size.

The statistical analysis of these data revealed the key biochemical differences that stratify sub-groups within the clinically relevant parameters. We have shown that the blister fluid proteome could be used to assist the classification of paediatric burn wounds by different burn depths and prognose time to re-epithelialisation at an earlier stage. Interestingly differences in the application of first aid may also be reflected in the burn wound protein profile. This may help deepen our understanding of the biological effect of first aid treatment.


Biography:

Tuo Zang is a PhD student of the Tissue Repair and Translational Physiology Program (TRTP) at the Institute of Health and Biomedical Innovation (IHBI) at the Queensland University of Technology. His research has focused on proteomics and metabolomics, especially quantitative proteomics analysis of clinical samples. His PhD study characterised blister fluid from paediatric burn injury using LC-MS/MS based proteomics and NMR based metabolomics.

How Bad is My Child’s Burn? Differences in Blister Fluid Biochemistry to Assess Paediatric Burn Wounds

Mr Tuo Zang1,2,3, Dr Leila Cuttle1,2,4, Dr Daniel Broszczak1,2,5, Dr James Broadbent1,2,6, Dr Catherine Tanzer1,2,4, Dr Tony Parker1,2

1Tissue Repair and Translational Physiology Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Australia, 2School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Australia, 3Wound Management Innovation Co-operative Research Centre, Brisbane, Australia, 4Centre for Children’s Burns and Trauma Research, Queensland University of Technology, Institute of Health and Biomedical Innovation at the Centre for Children’s Health Research, South Brisbane, Australia, 5School of Science, Faculty of Health Sciences, Australian Catholic University, Banyo, Australia, 6Sciex P/L, Brisbane, Australia

Abstract:

Burn blister fluid analysis provides an opportunity to non-invasively investigate the biology of the initial response to burn injury; uncover novel diagnostics or prognostics to assist in clinical decision making and enable the identification of new therapeutic approaches to enhance healing.

We performed a proteomic analysis of 87 paediatric burn blister fluid samples using liquid chromatography – tandem mass spectrometry with SWATH (data independent) acquisition, which allows for the large scale relative quantification of multiple proteins between samples. The blister fluid proteomes of all samples were compared to the key clinical features of burn depth classification and time-to-reepithelialisation. Both of these clinical parameters are critical for enabling accurate clinical decisions regarding early burn treatment options.

Subsequent analyses revealed significant differences in the biochemistry associated with both burn depth and time-to-reepithelialisation. Interestingly, the protein profiles provided evidence of potential clinical misclassification of some burn wounds examined in this study. Importantly, this highlights the utility of diagnostic markers for burn depth and / or prognostic markers of time-to-reepithelialisation. While full-thickness burns are often grafted and superficial-partial thickness burns are often not, it is more difficult to determine if deep-partial thickness burns should be grafted or not. Thus, utilisation of the results of this study could translate to aid in clinical decision making. Overall, this study provides new insights into the early stages of burn wound biology in children and may help with the development of diagnostic or prognostic tools to assist with clinical decisions regarding burn treatment options.


Biography:

Dr Tony Parker is the program leader of the Tissue Repair and Translational Physiology (TRTP) Program at the Institute of Health and Biomedical Innovation (IHBI) at the Queensland University of Technology. He is also a Senior Lecturer within the School of Biomedical Sciences at QUT. Dr Parker’s research has focused on the biochemical implications of musculoskeletal and skin tissue injury and recovery processes. In particular his team has utilized 2D and 3D in vitro skin wound models and developed the workflows required for the proteomic and metabolomic investigation of wound and tissue fluids. To achieve this he played a key role in the establishment of the world class proteomics and metabolomics capability at QUTs Central Analytical Research Facility (CARF). Dr Parker’s team also has an interest in the health promoting effects of physical activity wherein his team found that moderate levels of exercise in healthy individuals induces the systemic release of the neuroprotective / neurorepairative protein known as the LG3 peptide. This finding may have implications for stroke prevention and rehabilitation and his team is currently exploring the effect of exercise on the release of circulating LG3 peptide in a small clinical cohort.

Enhancing Clinical handover by using Burn External Referral form in a burn centre

Ms Tze Wing Wong1

1Hong Kong Hospital Authority, Burns Centre, Prince of Wales Hospital, Hong Kong

Abstract:

Background:Burns Centre, Prince of Wales Hospital is one of referral center for extensive burn cases in Hong Kong. It develops the coordination of burn service with their supporting network of Burn Facilities. Also, burn cases of cross-territory and cross center are consulted and transferred to the Centre for management. Therefore, a Burn External Referral form was designed to enable comprehensive clinical handover.

Objectives:

The establishment of Burn External Referral form enhances:

  1. Essential informations are received before burn patients transfer to Burns Centre.
  2. Guidance provides to healthcare professionals of other hospitals for managing acute burn stage.
  3. Referred burn patients receive acute assessment and management safely before transfer.

Methodology:A comprehensive Burn External Referral form was established. Essential and mandatory items for acute burn assessment were included, for example: Burn distribution, fluid challenge with formula, airway assessment, limbs circulation assessment, chemical burn treatment and treatment given prior transfer, etc. Briefing sessions were conducted to surgeons and nurses on the workflow of using the form. And auditing on compliance of Burn External Referral form was evaluated by using checklist.

Result:During Jun. 2016 to Jun. 2017, total 49 nos. of burn referred cases were received and 49 nos. of Burn External Referral forms were audited. Result showed only 87.8% sample forms documented the mechanism of burn injury. Only 47% sample forms showed correct assessment on area of burn injury.

Also, 98% and 93.9% indicated that burn patients’ airway and limb circulation had been examined before transferred. Only 75.5% sample forms had acknowledged the need for tetanus prophylaxis prescription.

Conclusion:The use of Burn External referral form provides guidance to healthcare professionals of referring hospitals for management and stabilization of burn patients. Essential informations are received for preparation for admission in Burns Centre. But, there still need for improvement in evaluating the area of burn injury which affects the treatment regime especially for extensive burn.

Reference:Australian Medical Association, 2006 Safe handover: safe patients. Guidance on clinical handover for clinicans and manager. Canberra ACT.


Biography:

Ms. Wong Tze Wing is Nurse Consultant in Burn and Plastic Specialty in Prince of Wales Hospital, Hong Kong Hospital Authority. She has extensive experience in burn management and regularly teaches and coaches nursing colleagues in Hong Kong. She also takes part in advances in burn care and is an EMBS course Nurse coordinator.

A simple method to colour balance digital photographs for remote assessment of tissue viability in acute burns

Mr Simon Chong1, Dr David Becker2

1Waikato Hospital, Hamilton, New Zealand, 2Waikato Hospital, Hamilton, New Zealand

Abstract:

Key to the practice of plastic surgery is the visual assessment of tissue perfusion via colour, most prominently when determining the depth of acute burns.

Digital imaging devices such as cell phones have provided a convenient means for remote assessment and referral of acute burns. However, most are optimised for use by inexpert photographers in non-medical applications. Accurate reproduction of colour is thus often compromised by environmental lighting, inconsistent use of flash illumination, and software bias. Digital devices will automatically optimise image parameters based upon the content of the image. White and black objects will be recognised and preferentially used to determine exposure and colour balance.

These variables often combine to produce a colour-inaccurate image with limited diagnostic utility and potential for misdiagnosis and liability.

We hypothesise that the routine placement of a white cotton swab within the photographic field of an acute burn will permit standardisation of colour balance across a variety of digital imaging devices.

Standardized photographs of a variety of burns will be taken with different devices. Six images will be taken: flash and non-flash with no swab present, flash and non-flash with a standard white cotton swab placed within the image framing, and flash and non-flash with a standard Pantone white colour swatch within the image.

The accuracy of the swab white balancing could be assessed with quantitative analysis utilizing Adobe Photoshop. Specifically, the variances in each Red Green Blue (RGB) colour channel could then be expressed as a percentage variance from the swatch-containing images.


Biography:

David is a Plastic Surgery registrar. Upon graduating from Otago University with an MBChB and Bachelor of Medical Laboratory Science in 2014 he worked in Dunedin as a house officer and surgical registrar; both in General Surgery and Plastic Surgery while completing a Postgraduate Diploma of Surgical Anatomy. Currently he is working at Waikato Hospital in Hamilton New Zealand.

Secure image storage: A new hope

Miss Siobhan Connolly1, Ms Anne Darton1

1ACI Statewide Burn Injury Service, St Leonards, Australia

Abstract:

A long time ago in a burn unit far, far away the benefits of digital images were recognised. These benefits were harnessed for the good of the patients and clinicians; and not just for burns.

One of the major ongoing challenges however is how to securely capture, forward and store these images to protect patient privacy. A project is underway to address this in NSW.

Burn clinicians have forged the way by consulting on what is required and how best to address the issue. It was decided to create a smart device application to enable images to be taken, sent but not stored on the device. Consultation also included non-burn clinicians from areas such as wounds, dermatology and emergency.

The saga continues……


Biography:

Siobhan has been working as the Burn Prevention/Education Manager of the ACI Statewide Burn Injury Service since November 2005. Before this time she nursed in the Burn Unit at The Children’s Hospital at Westmead for 9 years.
She is Chair of the ANZBA Burn Prevention Committee and the current ANZBA Treasurer. She also teaches burn management in NSW, across Australia and internationally and has led the development of numerous resources for burn management and prevention.

Children and families’ experiences of adhering to scar treatments

Ms Jessica Killey1, Dr Megan Simons1,2, Professor Roy Kimble1,2, Dr Zephanie Tyack1

1Centre for Children’s Burns and Trauma Research, UQ, South Brisbane, Australia, 2Lady Cilento Children’s Hospital, South Brisbane, Australia

Abstract:

Background:Despite being complex and time-consuming, non-invasive scar treatments such as pressure garments and silicone gels are frequently used to manage scarring in children for months or years after a burn. However, little is known about young children and their families’ experiences of adhering to these treatments.

Methods:This study aimed to develop a grounded theory of child and caregiver perceptions of adherence to scar treatments that will inform a new adherence intervention. Semi-structured interviews were conducted with children 8 to 16 years and caregivers of children 0 to 16 years who attended the burns centre at Lady Cilento Children’s Hospital, Brisbane for non-invasive scar treatments. Theoretical sampling was used to develop codes and categories. Information regarding socio-demographics, health literacy, comorbidity and scar symptoms was collected.

Findings:Thirteen interviews were conducted (mostly in participant’s homes) with caregivers of children receiving scar treatments (n = 11) and children themselves (n = 2). Caregivers were mostly female (n = 10, 91%) and their median age was 35 years (IQR: 30 – 40 years). Children included a male (12 years) and female (7 years). Preliminary codes included the use of distraction techniques to support pressure garment wear in young children and caregiver perceptions of what it feels like to wear a pressure garment. Caregivers described how incorporating scar treatments into everyday routines facilitated adherence.

Discussion: Based on the development of a grounded theory, implications for a new adherence intervention that may be used by children with burns and their caregivers will be described.


Biography:

Jessica Killey is an occupational therapist who has previously worked with adults with burn injuries. She is currently completing her PhD which is focussed on optimising adherence to scar treatments in children with burns and their families.

Take Six video resource for parents of young children undergoing burn dressing changes

Ms Erin Brown1,2, Dr Alexandra De Young1, Prof Roy Kimble1,3, Dr Bronwyn Griffin1,4, Dr Koa Whittingham2,5, Prof Justin Kenardy2

1Centre For Children’s Burns & Trauma Research, South Brisbane, Australia, 2School of Psychology, The University of Queensland, St Lucia, Australia, 3Pegg Leditschke Children’s Burns Centre, Lady Cilento Children’s Hospital, South Brisbane, Australia, 4Queensland University of Technology, Brisbane, Australia, 5The Queensland Cerebral Palsy and Rehabilitation Research Centre, South Brisbane, Australia

Abstract:

Background. Previous work has identified associations between parental psychological distress and young child distress behavior during acute burn wound care. In order to assist parents with supporting their child during burn wound care, we have developed a short video resource (6 minutes in length) for parents of young children (1-6 years old) presenting for the first burn outpatient dressing change. The video provides parents with psychological support, and behavioral strategies to assist their child during wound care. The video will be presented during this presentation. In addition, a preliminary study was conducted to test the acceptability and potential efficacy of the video resource. Methods. Parents were recruited to 1) provide acceptability feedback regarding the resource, and 2) to compare pilot data (N=8) with a previously observed control group (N=87). The comparative data focused on parent and child behaviour during burn wound care, and ratings of child anxiety and pain. Results. Parents found the video acceptable in terms of delivery and content, and had high intentions and confidence for using the behavioral strategies. The pilot study found parents demonstrated marginally lower rates of distress-promoting behavior compared to a control group, t(11.9)=-1.77, p=.054. In addition, children demonstrated significantly lower rates of distress behavior compared to a control group, t(8.4)=-1.90, p=.044. Thirdly, nurse-reported child peak pain score was significantly lower compared to a control group, t(12.1)=-2.95, p=.001. Conclusions. Take Six is an acceptable resource, and preliminary data indicates it is effective for improving parent and child experiences of burn wound care.


Biography:

Erin Brown has recently completed her PhD with the Centre for Children’s Burns and Trauma Research, and The University of Queensland.

Procedural pain and parental posttraumatic stress: Consequences for young child burn wound re-epithelization

Ms Erin Brown1,2, Dr Alexandra De Young1, Prof Roy Kimble1,3, Prof Justin Kenardy2

1Centre For Children’s Burns & Trauma Research, South Brisbane, Australia, 2The University of Queensland, St Lucia, Australia, 3Pegg Leditschke Children’s Burns Centre, Lady Cilento Children’s Hospital, South Brisbane, Australia

Abstract:

Background. The influence of psychological stress on wound healing has been found in several laboratory and medical populations. While pain has been linked to healing previously, the role of a parent’s wound-related stress regarding pediatric wound re-epithelization has not been investigated. Methods. The current study measured several aspects of the child’s and parent’s experience at the child’s first burn dressing change, and investigated the impact on wound re-epithelization. Eighty-three families’ behaviors were observed by a researcher. Parents completed psychological distress measures (posttraumatic stress symptoms, guilt, procedural anxiety, general anxiety/depression symptoms). Parents and nurses reported the child’s pre-, peak and post-procedural pain and anxiety. Days until re-epithelization were estimated based on when the wound clinically assessed as re-epithelized and the child was discharged from the clinic or referred to scar management.

Results. Days to re-epithelization ranged from 3-35 days post-injury. Using hierarchical multiple regression analyses, injury severity (wound depth and size) and parent education were entered as control variables. Control variables accounted for 37% of the variance, F(3,79)=15.26, p<.001. Other sample characteristics did not influence re-epithelization. Child peak procedural pain and child distress significantly influenced re-epithelization, ∆Fs(3,78)>5.10, ps<.027, ∆R2s>.04. One point increases accounted for 0.47 day and 0.99 day delays in re-epithelization, respectively. Parental PTSS and guilt also significantly influenced re-epithelization, ∆Fs(3,74)>4.08, ps<.047, ∆R2s>.03. One point increases accounted for 1.43 and 0.28 day delays in re-epithelization, respectively.

Conclusions. The findings indicate parent-reports of child procedural pain, parental traumatic stress and guilt are an important yet undervalued influences on paediatric re-epithelization.


Biography:

Erin Brown has recently completed her PhD with the Centre of Children’s Burns and Trauma Research, and the University of Queensland.

About ANZBA

ANZBA is a not for profit organisation and the peak body for health professionals responsible for the care of the burn injured in Australia and New Zealand. ANZBA encourages higher standards of care through education, performance monitoring and research.

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