Feeder layer- and animal product-free culture of neonatal foreskin keratinocytes: improved performance, usability, quality and safety

Since 1987, keratinocytes have been cultured at the Queen Astrid Military Hospital. These keratinocytes have been used routinely as auto and allografts on more than 1,000 patients, primarily to accelerate the healing of burns and chronic wounds. Initially the method of Rheinwald and Green was used to prepare cultured epithelial autografts, starting from skin samples from burn patients and using animal-derived feeder layers and media containing animal-derived products. More recently we systematically optimised our production system to accommodate scientific advances and legal changes. An important step was the removal of the mouse fibroblast feeder layer from the cell culture system. Thereafter we introduced neonatal foreskin keratinocytes (NFK) as source of cultured epithelial allografts, which significantly increased the consistency and the reliability of our cell production. NFK master and working cell banks were established, which were extensively screened and characterised. An ISO 9001 certified Quality Management System (QMS) governs all aspects of testing, validation and traceability. Finally, as far as possible, animal components were systematically removed from the cell culture environment. Today, quality controlled allograft production batches are routine and, due to efficient cryopreservation, stocks are created for off-the-shelf use. These optimisations have significantly increased the performance, usability, quality and safety of our allografts. This paper describes, in detail, our current cryopreserved allograft production process

Burn injury leads to increased long-term susceptibility to respiratory infection in both mouse models and population studies

Background: Burn injury initiates an acute inflammatory response that subsequently drives wound repair. However, acute disruption to the immune response is also common, leading to susceptibility to sepsis and increased morbidity and mortality. Despite increased understanding of the impact of burn injury on the immune system in the acute phase, little is known about longterm consequences of burn injury on immune function. This study was established to determine whether burn injury has long-term clinical impacts on patients' immune responses. Methods: Using a population-based retrospective longitudinal study and linked hospital morbidity and death data from Western Australia, comparative rates of hospitalisation for respiratory infections in burn patients and a non-injured comparator cohort were assessed. In addition, a mouse model of non-severe burn injury was also used in which viral respiratory infection was induced at 4 weeks post-injury using a mouse modified version of the Influenza A virus (H3NN; A/mem/71-a). Results and conclusions: The burn injured cohort contained 14893 adult patients from 1980-2012 after removal of those patients with evidence of smoke inhalation or injury to the respiratory tract. During the study follow-up study a total of 2,884 and 2,625 respiratory infection hospital admissions for the burn and uninjured cohorts, respectively, were identified. After adjusting for covariates, the burn cohort experienced significantly elevated admission rates for influenza and viral pneumonia (IRR, 95%CI: 1.73, 1.27-2.36), bacterial pneumonia (IRR, 95%CI: 2.05, 1.85-2.27) and for other types of upper and lower respiratory infections (IRR, 95% CI: 2.38, 2.09-2.71). In the mouse study an increased viral titre was observed after burn injury, accompanied by a reduced CD8 response and increased NK and NKT cells in the draining lymph nodes. This data suggests burn patients are at long-term increased risk of infection due to sustained modulation of the immune response

Defining family business: a closer look at definitional heterogeneity

Researchers have used a myriad of different definitions in seeking to explain the heterogeneity of family firms and their unique behavior; however, no widely-accepted definition exists today. Definitional clarity in any field is essential to provide (a) the basis for the analysis of performance both spatially and temporally and (b) the foundation upon which theories, frameworks and models are developed. We provide a comprehensive analysis of prior research and identify and classify 82 definitions of family business. We then review and evaluate five key theoretical perspectives in family business to identify how these have shaped and informed the definitions employed in the field and duly explain family firm heterogeneity. Finally, we provide a conceptual diagram to inform the choice of definition in different research settings

Facial burns in children: A series analysis with implications for resuscitation and forensic odontology

This study comprises a continuous (1981-1995) unselected series of all children who died from thermal injuries in the State of Queensland, Australia. One hundred and six children, so identified, died from incineration (35 per cent), respiratory burns with smoke or carbon monoxide inhalation (33 per cent), body surface area burns comprising greater than 60 per cent (9 per cent) and electrocution (20 per cent). The burn fatality rate was 0.98 per hundred thousand children (0-14 years) per year, with no secular trend and, specifically, no reduction in the annual rate of such fatalities. Eighty-two children (49 males) had concomitant facial injuries, both thermal and nonthermal; of whom 55 per cent were under the age of five years. Sixty (73 per cent) child burn victims died in house fires. Forensic odontology is important in confirming the age of such victims in single incinerations but is of limited value when larger numbers of children are incinerated, because of the relative lack of dental restorations in the infant and pre-school age group. Of the 82 children with facial and airway injuries, 12 per cent had only mild or superficial facial damage and only seven (8 per cent) were alive or resuscitatable at the time of rescue from the conflagration or burning injury. Child deaths from burns contributed an annual loss rate of 506 years of potential life lost (YPLL) in a population of 3 million of whom 21.5 per cent were children under the age of 15 years. Airway management and resuscitation, in the context of managing surviving burn victims of any age with facial injuries, pose special difficulties. Inhalational burns (smoke and the gases of conflagration) result in a mortality greater than 60 per cent. Although 81 per cent of children showed evidence of airway obstruction, analysis of current data indicates that a maximum of 8 per cent could have survived with airway maintenance and protection. Inhalational burns (to both upper and lower airways) grossly reduce survivability. Primary prevention would seem vital and thus remains a major challenge to reduce the incidence of such deaths. Some strategies include advocacy to promote the compulsory installation of smoke alarms, family drills to practise escape and the teaching of 'first aid for all'