p62 overexpression induces TDP-43 cytoplasmic mislocalisation, aggregation and cleavage and neuronal death.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) that exist on a spectrum of neurodegenerative disease. A hallmark of pathology is cytoplasmic TDP-43 aggregates within neurons, observed in 97% of ALS cases and ~ 50% of FTLD cases. This mislocalisation from the nucleus into the cytoplasm and TDP-43 cleavage are associated with pathology, however, the drivers of these changes are unknown. p62 is invariably also present within these aggregates. We show that p62 overexpression causes TDP-43 mislocalisation into cytoplasmic aggregates, and aberrant TDP-43 cleavage that was dependent on both the PB1 and ubiquitin-associated (UBA) domains of p62. We further show that p62 overexpression induces neuron death. We found that stressors (proteasome inhibition and arsenic) increased p62 expression and that this shifted the nuclear:cytoplasmic TDP-43 ratio. Overall, our study suggests that environmental factors that increase p62 may thereby contribute to TDP-43 pathology in ALS and FTLD.

ALS-associated TBK1 variant p.G175S is defective in phosphorylation of p62 and impacts TBK1-mediated signalling and TDP-43 autophagic degradation.

Mutations affecting SQSTM1 coding for p62 and TANK-Binding Kinase 1 (TBK1) have been implicated in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). TBK1 is a serine-threonine kinase that regulates p62's activity as an autophagy receptor via phosphorylation and also has roles in neuroinflammatory signalling pathways. The mechanisms underlying ALS and FTLD pathogenesis as a result of TBK1 mutations are incompletely understood, however, loss of TBK1 function can lead to dysregulated autophagy and mitophagy. Here, we report that an ALS-associated TBK1 variant affecting the kinase domain, p.G175S, is defective in phosphorylation of p62 at Ser-403, a modification critical for regulating its ubiquitin-binding function, as well as downstream phosphorylation at Ser-349. Consistent with these findings, expression of p.G175S TBK1 was associated with decreased induction of autophagy compared to wild type and reduced degradation of the ALS-linked protein TDP-43. Expression of wild type TBK1 increased NF-κB signalling ~300 fold in comparison to empty vector cells, whereas p.G175S TBK1 was unable to promote NF-κB signalling above levels observed in empty vector transfected cells. We also noted a hitherto unknown role for TBK1 as a suppressor of oxidative stress (Nrf2) signalling and show that p.G175S TBK1 expressing cells lose this inhibitory function. Our data suggest that TBK1 ALS mutations may broadly impair p62-mediated cell signalling, which ultimately may reduce neuronal survival, in addition TDP-43 was not efficiently degraded, together these effects may contribute to TBK1 mutation associated ALS and FTLD pathogenesis.

An FTLD-associated SQSTM1 variant impacts Nrf2 and NF-κB signalling and is associated with reduced phosphorylation of p62.

Elevated oxidative stress has been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS) and Frontotemporal Lobar Degeneration (FTLD). In response to oxidative stress, the Nrf2 transcription factor activates protective antioxidant genes. A critical regulator of Nrf2 is the inhibitory protein Keap1, which mediates Nrf2 degradation. In response to cellular stress an interaction between Keap1 and SQSTM1/p62 (p62), a signalling adaptor protein, allows for increased Nrf2 signalling as it escapes degradation. Mutations in SQSTM1 (encoding p62) are linked with ALS-FTLD. Previously, two ALS-FTLD-associated p62 mutant proteins within the Keap1 interacting region (KIR) of p62 were found to be associated with decreased Keap1-p62 binding and Nrf2 activation. Here we report that a non-KIR domain FTLD-associated variant of p62 (p.R110C), affecting a residue close to the N-terminal PB1 oligomerisation domain, also reduces Keap1-p62 binding in cellulo and thereby reduces Nrf2 activity in reporter assays. Further, we observed that expression of p.R110C increased NF-κB activation compared with wild type p62. Altered signalling appeared to be linked with reduced phosphorylation of p62 at Serine residues -349 and -403. Our results confirm that ALS-FTLD mutations affecting multiple domains of p62 result in a reduced stress response, suggesting that altered stress signalling may directly contribute to the pathology of some ALS-FTLD cases.

Targeted sequencing of DCSTAMP in familial Paget's disease of bone.

Paget's disease of bone (PDB) has a strong genetic component. Variants in SQSTM1 are found in up to 40% of patients with a family history of the disease, where a pattern of autosomal dominance with incomplete penetrance is apparent. By contrast, SQSTM1 variants are only found in up to 10% of patients with sporadic disease. It has been hypothesised that the remaining genetic susceptibility to PDB, particularly in familial cases, could be explained by rare genetic variants in loci previously identified by Genome Wide Association Studies. It is likely that polygenic factors are involved in many individuals. In this study we utilised whole exome sequencing to investigate predisposing genetic factors in an unsolved PDB kindred and identified a c.1189C > T p.L397F variant in DC-STAMP, also known as TM7SF4, that co-segregated with disease. DCSTAMP was identified as a gene of interest in PDB following Genome Wide Association Studies and has been previously shown to play critical roles in osteoclast fusion. The variant we identified has also been reported in association with PDB in a French-Canadian cohort however the significance of this variant was inconclusive. Targeted screening of DCSTAMP in our familial cohort of PDB patients revealed an additional 8 variants; however we did not find a significant association between any of these, including p.L397F, with PDB. Osteoclastogenesis assays from the affected proband and his unaffected brother demonstrated an increase in osteoclast number and nucleation, consistent with the pagetic phenotype. In converse to other established Paget's associated genetic variations such as SQSTM1, TNFRSF11A and OPTN, expression of the mutant DC-STAMP protein attenuated the activation of transcription factors NFκB and AP-1 when exogenously expressed. We found that the p.L397F variant did not influence the subcellular localization of the protein. Based on these findings we conclude that genetic variation in DCSTAMP is not a significant predisposing factor in our specific cohort of PDB patients and the p.L397F variant is unlikely to be a contributing factor in PDB pathogenesis.

Do airline pilots and cabin crew have raised risks of melanoma and other skin cancers? Systematic review and meta-analysis.

BACKGROUND: Airline pilots and cabin crew are potentially exposed to hazardous ultraviolet and cosmic radiation, which may increase their risk of melanoma and other skin cancers. OBJECTIVES: To establish precise risks of melanoma and keratinocyte cancer (KC) for airline pilots and for cabin crew based on all studies published to date. METHODS: We searched MEDLINE, ISI Science Citation Index, Embase, SCOPUS and CINAHL to June 2018. All studies of melanoma and KC risk and mortality in airline pilots and cabin crew compared with the general population were eligible. Standardized incidence ratios (SIRs) and standardized mortality ratios (SMRs) were pooled using random effects models. RESULTS: From 5866 papers retrieved, we reviewed 44 full-text articles, of which 12 studies with data collected mostly between the 1970s and 1990s were eligible for inclusion. The pooled SIR (pSIR) for melanoma in pilots was 2.03 [95% confidence interval (CI) 1.71-2.40] and in cabin crew it was 2.12 (95% CI 1.71-2.62). For pilots, the pooled SMR for melanoma was 1.99 (95% CI 1.17-3.40) and for cabin crew it was 1.18 (95% CI 0.73-1.89). For KC, the pSIR was 1.86 (95% CI 1.54-2.25) in pilots and 1.97 (95% CI 1.25-2.96) in cabin crew. There was no evidence of study heterogeneity. CONCLUSIONS: The available evidence shows that airline pilots and cabin crew have about twice the risk of melanoma and other skin cancers than the general population, with pilots more likely to die from melanoma. However, most of the evidence was collected several decades ago and their relevance to contemporary levels of risk is uncertain.

A feasibility study for a triple-blind randomized controlled trial investigating the effects of oral isotretinoin on mood and quality of life in patients with acne vulgaris.

Isotretinoin is used in the treatment of severe acne vulgaris (AV), but has controversially been associated with depression and suicide. Large prospective studies have failed to translate this clinically. We undertook a feasibility study to investigate the parameters of a triple-blind, randomized controlled trial (RCT) assessing the effect of oral isotretinoin on quality of life (QoL) and mood in patients with AV. Patients meeting the inclusion criteria were randomized for 2 weeks to isotretinoin or doxycycline. Participants completed verified depression and QoL screening questionnaires at baseline and week 2. In total, 194 patients with AV were screened, with 48 meeting the inclusion criteria and 13 of these being willing to participate. The follow-up rate was 92% and questionnaire response rate was 96%. To our knowledge, this is the first study to demonstrate a successful design for a triple-blind RCT investigating the effects of isotretinoin on mood in patients with AV.

Fatty thymic involution uptake mimicking a thymic epithelial tumour in a 18F-FDG PET/CT study / Captura de involución tímica grasa imitando un tumor epitelial tímico en un estudio 18F-FDG PET/TC

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Electroconvulsive Therapy - What Do Patients Think Of Their Treatment?

BACKGROUND: The Regulation and Quality Improvement Authority (RQIA) monitors the administration of electroconvulsive therapy (ECT) in Northern Ireland (NI). As part of their inspection methodology RQIA wished to include feedback from ECT patients. The aim of this report is to summarise the opinions of ECT patients over a 1-year period and to compare their feedback about treatment with the standards of best practice, as defined by the Electroconvulsive Therapy Accreditation Service (ECTAS). METHOD: RQIA was granted permission to use the ECTAS patient questionnaire. The questionnaire was distributed to all the ECT clinics in NI and staff were requested to give them to patients who had received a course of ECT. RESULTS: A total of 42 individuals returned questionnaires, 24 females (57.1%) and 18 (42.9%) males. The response rate was 26%. Almost half of respondents were detained under the Mental Health (Northern Ireland) Order 1986 (n=19, 45.2%), with one third receiving ECT as a day patient (n=14, 33.3%). Respondents reported having detailed information about ECT, with ECTAS standards 4.2 and 4.3 being affirmed in over 80% of cases. Eighty percent of respondents (n=34) believed they benefited from ECT. CONCLUSION: The results are mainly favourable towards ECT. The majority felt they benefited from treatment.

Welfare indicators during broiler slaughtering.

1. The aim of this study was to identify the most relevant welfare indicators for unloading, lairage, stunning, killing and post-mortem inspection in a poultry slaughter plant. Different indicators were unloading duration, lairage time, environmental variables in the lairage facilities, shackling time and electrical variables used in the water bath. 2. Lairage time did not correlate strongly with dead on arrival. Heat stress was limited by means of ventilation systems, correct cage placement and appropriate stocking density per crate. The acceptable shackling period was about 30 s. 3. The presence of a corneal reflex showed that an animal was alive, while spontaneous wing flapping, spontaneous eye blinking and response to a painful stimulus were regarded as indicators of stunning efficiency. 4. It was concluded that the presence of recent traumatic injuries during the post-mortem inspection could be a valid means to establish whether corrective measures concerning the handling, transport and loading procedures should be taken.

Technical note: Protozoa-specific antibodies raised in sheep plasma bind to their target protozoa in the rumen.

Binding of IgG antibodies to Entodinium spp. in the rumen of sheep (Ovis aries) was investigated by adding IgG, purified from plasma, directly into the rumen. Plasma IgG was sourced from sheep that had or had not been immunized with a vaccine containing whole fixed Entodinium spp. cells. Ruminal fluid was sampled approximately 2 h after each antibody dosing. Binding of protozoa by a specific antibody was detected using an indirect fluorescent antibody test. An antibody titer in the ruminal fluid was determined by ELISA, and the concentration of ruminal fluid ammonia-N and ruminal pH were also determined. Entodinium spp. and total protozoa from IgG-infused sheep were enumerated by microscopic counts. Two-hourly additions of IgG maintained a low antibody titer in the rumen for 12 h and the binding of the antibody to the rumen protozoa was demonstrated. Increased ammonia-N concentrations and altered ruminal fluid pH patterns indicated that additional fermentation of protein was occurring in the rumen after addition of IgG. No reduction in numbers of Entodinium spp. was observed (P>0.05). Although binding of antibodies to protozoa has been demonstrated in the rumen, it is unclear how much cell death occurred. On the balance of probability, it would appear that the antibody was degraded or partially degraded, and the impact of this on protozoal populations and the measurement of a specific titer is also unclear.

The occurrence of giant fibres in different muscles of two chicken genotypes.

1. The occurrence of Giant Fibres (GF) in three muscles (Pectoralis major (PM), Iliotibialis lateralis and Semimembranosus) with different types of energy metabolism was studied in slow- and fast-growing chicken strains. 2. A total of 20 one-day-old Leghorn chicks (slow-growing) and 20 broiler (Ross 508) chicks (fast-growing) were reared to 100 and 45 d, respectively. 3. A small percentage of GF was seen in pre rigor muscle samples even at 3 min post mortem in both genotypes and in all muscle types studied. 4. From 3 min to 24 h post mortem GF increased both in Leghorn and broiler chickens but to a different extent according to muscle type and genotype. 5. The highest GF 24 h post mortem value was found in the PM muscles belonging to the fast-growing broiler line. 6. It was concluded that every type of muscle can develop GF, but this phenomenon is more evident in the PM especially in animals selected for increased growth rate.

Hot ash burns in the children of Western Australia: how and why they happen.

INTRODUCTION: Burns from hot ash are common in the paediatric population in Western Australia. Fifty children were admitted to the paediatric burn centre with hot ash contact burns to the feet in 2011 and 2012. It is important to examine the extent of the problem, seasonal variations, and identify those at risk to determine strategies for prevention campaigns. METHOD: Retrospective review of medical notes for all admissions to the paediatric burns unit was undertaken for 2011 and 2012. Data were collected for patient demographics, time, circumstance of injury, burn severity and treatment. RESULTS: Hot ash burns accounted for 8.6% of admissions but 16.1% of burns sustained in non-metro areas. Median age was just under 3 years, male or female. Median burn TBSA was 2%, and 44% of children required surgery. The burns were less common in summer, more common on non-school days and in children who were on camping trips away from home. DISCUSSION: Previous work has shown the value of targeted campaigns. The group for targeted prevention campaigns are the carers of very young children who go camping. Information distributed at camping shows and stores about the principles of campfire safety would reach the people at risk.

In the media: Burns as a method of assault.

AIM: The aims of this study were to determine whether a change occurred in the pattern of assault burn injury cases hospitalised to the adult state burns unit, Western Australia, from 2004 to mid-year of 2012, and to compare patient and burn characteristics of adult assault burns with those admitted for unintentional burns. METHODS: Study data were obtained from the Royal Perth Hospital (RPH) Burns Minimum Dataset (BMDS). Aggregated data of unintentional burn admissions during the same period were provided by the BMDS data manager to enable comparisons with assault burn patients. RESULTS: Assault burn admissions during 2004-2012 accounted for approximately 1% of all adult burn hospitalisations. All assault victims were burned by either thermal or scald agents. A high rate of intubation (24%) and ICU admission (1 in 3 cases) was observed in the fire assault group. The six assault cases undergoing intubation were severe burns, median TBSA 50%, most commonly affecting the face, head and torso, half of these cases had inhalational injuries and also required escharotomies. Comparison of admissions by calendar period showed no statistically significant differences in demographic, burn cause or TBSA%. However, statistically significant differences were found for pre-morbid psychiatric history (15% vs. 58%, p=0.025) and concomitant fractures or dislocations (46% vs. 2%), p=0.011). CONCLUSIONS: While the proportion of assault burn admissions per total burn admissions steadily increased from 0.4% in 2009 to 1.5% in mid-2012, this proportion did not exceed that peak level observed of 2.1% for 2004.

Sweet's syndrome mimicking alkali burn: a clinical conundrum.

INTRODUCTION: Sweet's syndrome or acute febrile neutrophilic dermatosis presents most commonly on the hands, upper extremities and face. The disease is of clinical relevance to surgeons as it could mimic an infective aetiology but debridement negates such wounds. CASE: A 34-year-old man was referred from a peripheral hospital with suspected infected alkali burn to the hands. A builder by profession, he had been working outdoors with possible exposure to cement-concrete mix, 5 days previously. At presentation, the dorsal aspect of the thenar eminence appeared erythematous and oedematous, with pustules and blisters with central ulcerations. Haematological investigation revealed a neutrophilic leucocytosis and raised CRP. On the second day of admission the patient became febrile. He was treated with analgesia, IV Tazocin (Pipperacillin and Tazobactam) and the wounds were surgically debrided and covered using autologous cell therapy via the Recell kit. Two days following surgery, microbiology of wound swabs, tissue samples and blood cultures yielded no growth. The wound was noted to be extending beyond the zone of injury and a new area of erythema was evident on the neck. A diagnosis of idiopathic acral Sweet's syndrome was confirmed when histopathological investigation showed a moderate inflammatory cell infiltrate in the dermis. A rapid response to oral corticosteroids was clinically evident after 48 h and the lesions were completely healed at 4 weeks follow-up. CONCLUSION: We recommend thorough clinical history and examination, systematic wound review, tissue biopsy and culture in conjunction with dermatology opinion in cases of suspected Sweet's syndrome. Surgical debridement should be avoided as it has the potential to negate such wounds secondary to pathergy phenomenon.

A prospective randomised clinical pilot study to compare the effectiveness of Biobrane® synthetic wound dressing, with or without autologous cell suspension, to the local standard treatment regimen in paediatric scald injuries.

BACKGROUND: Scald is the most common cause of burn in children in Australia. The time taken by the burn wound to heal impacts on scar outcome. Commonly scald injuries are treated conservatively; in our unit the practice is that if healing does not occur within 10 days, surgery is used to aid healing with the aim of improving scar outcome. This randomised controlled pilot study compares early treatment regimens to facilitate tissue salvage and reduce the incidence of definitive surgery at 10 days following scald injury. METHODS: All paediatric patients with partial thickness scald injury were clinically assessed between July 1, 2009 and June 30, 2010. A burn of 2% TBSAB or more and deemed not to heal within 10 days, were considered for the trial. These patients were randomised to one of three treatment arms: the local standard treatment (Intrasite™, Acticoat™ and Duoderm(®) dressings every 2-3 days) with surgery at 10 days, Biobrane(®) only or Biobrane(®) and autologous cell suspension using the ReCell(®) kit. The primary outcome was surgery performed after 10 days; secondary outcomes were rates of healing, pain experienced, and scar outcomes. RESULTS: 15% of scald presentations in the 12 month period met the eligibility criteria. 13 patients were recruited into the pilot study; early intervention was associated with a decreased time to healing with fewer dressing changes, less pain and better scar outcomes. CONCLUSION: Investment of surgical resources in the acute stages within 4 days of injury saved on nursing time, dressing, analgesic and scar management costs.

An assessment of burn injury hospitalisations of adolescents and young adults in Western Australia, 1983-2008.

BACKGROUND: This study presents a 26-year epidemiological assessment of burn injury hospitalisations for people 15-29 years of age in Western Australia. METHODS: Linked hospital morbidity and death data for all persons hospitalised with an index burn injury in Western Australia for the period 1983-2008 were analyzed. Annual age-specific incidence rates were estimated. Poisson regression analyses were used to estimate temporal trends in hospital admissions. RESULTS: There were 6404 burn hospital admissions of which 76% were male. Males had hospitalisation rates 3.0 times that of females (95%CI: 2.8-3.2) and Aboriginal people had rates 2.3 times (95%CI: 2.1-2.5) that of non-Aboriginal persons. Hospitalisations for burn injury declined by 42% (95%CI: 35-47) for males and 21% (95%CI: 6-33) for females. Hospitalisations declined by 53% (95%CI: 35-63) for Aboriginal people, and by 35% (95%CI: 29-41) for non-Aboriginal people. Significant reductions were observed for flame and electrical burn hospitalisations. The major causes of burns in males were exposure to controlled fires and ignition of inflammable materials, with scalds the predominant cause of burn in females. CONCLUSIONS: Downward trends in burn injury hospitalisations for both males and females 15-29 years of age were observed; however, males and Aboriginal persons have significantly elevated hospitalisation rates.

Demonstration of the use of the ICF framework in detailing complex functional deficits after major burn.

Burns can result in long term impairments, activity limitations and participation restrictions in a patients' life. The focus of current surgeries and therapy is to improve body functions and structures. However, often this does not translate to an improvement in activity and participation for the patient. Improvement in activity and participation is the ultimate goal of all therapy to enhance patient's quality of life. The incorporation of assessment measures at all levels of the International Classification of Functioning, Disability and Health (ICF) can assist in a holistic, patient centred approach to identify the complex impairments that impact on activity and participation, with a view to appropriately targeting future therapeutic interventions. This paper presents an example case of how implementing measures at all levels of the ICF can improve our understanding of a patient's body functions and structures, activity and participation. A number of the outcome measures utilised in this study are novel in the burns population, such that video footage supplements the methodology where relevant.

Methamphetamine laboratory-related burns in Western Australia--why the explosion?

INTRODUCTION: With increasing numbers of illicit drug users in both urban and rural communities, users and producers are becoming increasingly enterprising in their sourcing of mind altering drugs. An example of this is the 'amateur' production of methamphetamine in domestic dwellings. We describe the mechanism of burn seen in methamphetamine production, the pattern of clinical injury, and the difficulties in treating these patients. METHODS: A 12 month retrospective study of five patient groups presenting to our burn service with injuries following methamphetamine laboratory explosion. RESULTS: Out of five patient groups we have treated 9 individual patients (with one patient presenting on two different occasions) with burns following methamphetamine laboratory explosion. All patients were male and required hospital admission. The cause of the explosive injury was initially reported as barbeque or oven related, assault, or accident in all patients. Two patients (in separate events) required intubation for associated inhalation injury. Burn size varied from 1% to 40% BSA. 7 patients required surgical debridement and skin grafting. Injury type was thermal and chemical. All patients had difficult follow-up due to low levels of clinic attendance. CONCLUSION: Methamphetamine laboratory explosion burns are difficult injuries from the start. Invariably the true circumstances surrounding the injury are not clear, and clinicians should be suspicious of a meth lab explosion in suspect individuals with burns plus airway injury. Patient management is complex and often requires substantial analgesic and anxiolytic medication in conjunction with clinical psychology and psychiatry as an inpatient.