The novel epoxy-tiglianes, EBC-46 and ‘lesser activity’ EBC-211, are sourced from seeds of the Fountain’s Blushwood Tree (Fontainea picrosperma), indigenous to Queensland Tropical Rainforest. Australian biotechnology company, QBiotics Ltd.,has demonstrated that EBC-46 stimulates exceptional dermal wound healing responses in vivo, following cancer treatment and tumour destruction in domesticated animals. Consequently, QBiotics is developing EBC-46 as a veterinary anti-cancer pharmaceutical and performing human clinical trials. However, little is known on how EBC-46 induces its exceptional healing effects, manifested as accelerated wound re-epithelialisation, closure and reduced scarring. This study aimed to elucidate how EBC-46 and EBC-211 mediates these exceptional wound healing effects in vitro, through analysis of HaCaT keratinocyte and dermal fibroblast/myofibroblast genotypic and phenotypic responses, following epoxytigliane treatment (0.001-100μg/ml). A number of key wound healing responses were assessed, including proliferation, cell cycle progression, scratch wound repopulation; and transforming growth factor-β1 (TGF-β1)-driven, fibroblastmyofibroblast differentiation. Studies demonstrated that both EBC-46 and EBC-211 induced fibroblast and HaCaT cytotoxicity at 100μg/ml. EBC-46 and EBC-211 (0.001-10μg/ml) significantly retarded fibroblast proliferation and delayed S/G2 cell cycle transition, but exerted no significant effects on fibroblast migratory responses. Although EBC-46 had no effects on α-smooth muscle actin (α-SMA) expression, stress fibre organization and myofibroblast formation (0.001-0.01μg/ml and 1-10μg/ml), EBC-46 significantly inhibited α-SMA expression and stress fibre formation at 0.1μg/ml, with cells retaining normal fibroblast morphologies. EBC-211 induced similar effects at 10μg/ml Both EBC-46 and EBC-211 (0.001-10μg/ml) stimulated significant HaCaT proliferation, G1/S and S/G2 cell cycle transitions; and accelerated scratch wound repopulation, even with mitomycin C. Microarray analysis and protein level validation, identified numerous differentially expressed genes in epoxy-tiglianetreated, HaCaTs. Up-regulated genes included certain keratins and others associated with promoting cell cycle progression, proliferation and migration. Down-regulated genes included other keratins and genes associated with inhibiting cell cycle progression and proliferation, including certain cytokines and chemokines. This study has provided evidence to explain the enhanced re-epithelialisation and reduced scarring responses observed in epoxy-tigliane-treated skin. Furthermore, it highlights the potential of epoxy-tiglianes as novel therapeutics for impaired dermal wound healing and excessive scarring situations.

No description available.

No description available.

Mesenchymal stem cells (MSC) have been used clinically to treat a range of conditions, and have been shown to enhance skin wound healing in vivo. There is evidence suggesting that the beneficial effects of MSC transplantation may be mediated more through their paracrine activity than through their differentiation into mature cell types. This thesis has sought to examine the effects of MSC secreted factors on dermal fibroblasts, keratinocytes, and endothelial cells, using an in vitro wound healing model. The results presented demonstrate that mesenchymal stem cell-conditioned culture medium (MSC-CM) enhances wound closure by dermal fibroblasts and keratinocytes through increased skin cell migration. MSC-CM was similarly shown to enhance wound closure by endothelial cells. The contents of MSC-CM were analysed by immunological techniques and mass spectrometry to reveal targets for further investigation. These included cytokines, notably interleukin-6 (IL-6), interleukin-8 (IL-8) and transforming growth factor-beta one (TGF-β1); and extracellular matrix (ECM) components, notably collagen type I, fibronectin and decorin. Examining these factors individually or in combination with each other or MSC-CM demonstrated differential and interdependent effects. Fibronectin was generally stimulatory to wound closure, whilst decorin was inhibitory. The cytokines studied had variable effects, e.g. TGF-β1 was stimulatory to fibroblast migration in combination with collagen type I, elicited no effect in combination with fibronectin, and was inhibitory to keratinocyte migration in both conditions. This study has thereby progressed our understanding of how MSC secreted factors influence skin wound healing and suggests that further investigation of MSC activity is needed in order to inform the eventual clinical use of MSC to treat severe or chronic skin wounds. With further work, MSC-related therapy, using MSC transplantation or the delivery of MSC secreted products, might provide a preferable alternative to current treatment strategies for these wounds, either in terms of patient benefit or clinical cost reduction.

Wet-work exposure has been recognized as a main risk factor for developing Irritant contact dermatitis of the hand. Duration and frequency of wet-work exposure are the key risk determinants, but there is still no reliable objective method to assess these factors in the workplace. The current study aimed to further validate a newly invented wet-work exposure monitor (WWEM) among four high risk occupational groups of hairdressers, florists, caterers and nurses. The WWEM, which consists of two similar thermocouples provides a real-time indication of the wetness of the finger by measuring the temperature changes when the finger is wet and subsequent cooling when the water evaporates from the skin. A suitable data analysis routine was constructed and direct observation was employed as the gold standard. Receiver Operating Characteristic (ROC) analysis was used to compare the results. In laboratory experiments, the most suitable wet-event threshold value, as a criterion to distinguish the wet and dry exposure periods was identified as 1.6°C (Mean+5SD of the “dry” data). Using this value sensitivity and specificity were 70% and 78%, respectively. An area under the curve (AUC) of 0.78 demonstrated a “high” accuracy for WWEM in lab experiments. Field testing in real occupational fields identified varying degrees of user acceptability and different wet-event threshold values to produce maximal sensitivity and specificity of the instrument. Among hairdressers, the WWEM showed a “moderate” accuracy at the threshold value of 1.37°C with an AUC of 0.62. For florists, the most suitable threshold value in the range of examined thresholds was 1.14°C with a “good” accuracy. Among a cohort of caterers, a threshold value of 1.6°C demonstrated a “hi gh” accuracy with an AUC of 0.72. The WWEM demonstrates the minimum value of the largest AUC for nurses. This is the minimum amount of accuracy among the four occupational groups. The figure is 0.52, achieved at a threshold value of 2.5°C. The WWEM enables the frequency and duration of wet-work exposure to be assessed in an objective manner rather than the current costly and unreliable subjective methods of direct observation and questionnaire. The results indicate that the WWEM has good sensitivity and high specificity in detecting exposure to wet-work. This device provides new experimental data on wetwork exposure and may be used in future as an educational tool to highlight the importance of wet-work exposure to both employees and employers.

Introducción: La hipótesis de cierre intermitente del tubo neural justificaría laasociación de aplasia cutis congénita de cuero cabelludo (ACCCC) con raquisquisis anivel lumbo-sacro sin defectos de la piel suprayacente a ese nivel, hecho del que noexisten precedentes en la literatura.Objetivos: 1.- Estudio de 35 casos diagnosticados en el Hospital Clínico Universitariode Valencia entre Septiembre de 1993 y Diciembre de 2004. 2.- Análisis estadístico deasociaciones entre las distintas características de los pacientes. 3.- Valorar si la ACCCCes marcador de defectos de cierre del tubo neural y/o de otros defectos de congénitos,especialmente de la línea media, tanto subyacentes como a distancia. 4.- Nuevoprotocolo diagnóstico. 5.- Dermatoscopia. 6.- Nueva clasificación de la ACCC.Conclusiones: Existe relación entre presencia de collarete alrededor de la ACCCC yraquisquisis o cranio-raquisquisis. Este collarete es marcador de raquisquisis oculta adistancia. La forma hipertrófica de ACCCC asocia más raquisquisis a distancia que lasformas atrófica y membranosa. Las lesiones de mayor tamaño tienen mayor tendencia ala agregación familiar. El tipo membranoso es el que menos se hereda y el atrófico elque más. La presencia de ACCC fuera del vértex aumenta el riesgo de lesionessubyacentes. La forma membranosa suele localizarse en vértex. Las lesiones máspequeñas asocian con mayor frecuencia lesiones subyacentes. El grupo 1 de Friedentiene las lesiones de menor tamaño. Existe relación significativa entre tipo clínico y áreade la lesión (tipo atrófico más grandes). La presencia de collarete es más frecuente enlos casos con alteraciones de la línea media. El grupo 1 de Frieden tienen menorposibilidad de defecto subyacente que los grupos 2 y 3. La forma membranosa no asociadefectos subyacentes. El tipo clínico atrófico es el que más se relaciona con presenciade otras malformaciones congénitas fuera de la línea media. Las formas membranosa ehipertrófica pueden asimilarse al grupo 1 de Frieden. No puede afirmarse con todaseguridad que la presencia de ACCCC en sí misma es marcador de raquisquisis o lo essolamente cuando aparece asociada a otros defectos de la línea media. Hay mayorprobabilidad de encontrar raquisquisis en los pacientes con ACCCC que en la poblaciónen general. Además esta raquisquisis aparece a distancia sin que exista alteracióncutánea suprayacente en la mayoría de pacientes. El protocolo diagnóstico debe incluirtécnicas para investigar la posible presencia de raquisquisis oculta a distancia u otrosdefectos ocultos de la línea media a distancia. La dermatoscopia puede ser útil en eldiagnóstico de la ACCCC y otras alteraciones congénitas con signo del collarete. Lanueva clasificación propuesta según tres tipos clínicos (membranoso, atrófico ehipertrófico), facilita el estudio de la ACCCC y mejora en algunos aspectos laclasificación de Frieden. / Introduction: The hypothesis of intermittent pattern of neural tube closure could justifythe association of congenital aplasia cutis of the scalp (CACS) with lumbo-sacralrachischisis without cutaneous defects over the rachischisis. To our knowledge, thisassociation has never been reported so far.Objetives: 1.- Study of 35 patients with CACS from Hospital Clínico Universitario inValencia, Spain, between September 1993 and December 2004. 2.- Statistical analysisof the associations among the different features of the patients. 3.- Verify if CACS is acutaneous marker of neural tube closure defects and/or other congenital defects,specially of the middle line, underlying the cutaneous defect or distant to it. 4.- Newdiagnostic protocol. 5.- Dermoscopy. 6.- New classification for CACS.Conclusions: The presence of hair collars in CACS is related to rachischisis or craniorachischisis.This hair collar is a marker for distant occult rachischisis. HypertrophicCACS is more frequently associated to distant rachischisis than atrophic or membranousCACS. Bigger lesions tend to be more hereditary. Membranous CACS is the leasthereditary type and atrophic CACS is the most. When CACS is not located on thevertex the risk of underlying lesions is increased. Membranous CACS is generallylocated on the vertex. The smallest lesions associate more frequently underlying defects.Frieden type 1 associates the smallest lesions. There is statistically significativerelationship between the clinical type of CACS and the area of the lesion (atrophicCACS are bigger). The presence of hair collar is more frequent when there arealterations in the middle line. Frieden group 1 associates less often underlying defectscompared to groups 2 and 3. Membranous CACS does not associate underlying defects.Atrophic CACS is the type more often associated to other congenital malformations outof the middle line. Membranous and hypertrophic types constitute Frieden group 1. Wecan not confirm that the presence of CACS by itself is a marker of rachischisis or if itneeds to be associated with other middle line defects. There is a higher probability ofrachischisis in patients with CACS than in the rest of the general population. Andbesides this is a distant rachischisis with no cutaneous defects over the rachischisis inmost of the patients. The diagnostic protocol should include tests in order to detectpossible distal occult rachischisis or other distant occult middle line defects.Dermoscopy can be useful for the diagnosis of CACS and other congenital defectsassociating hair collar sign. The new proposal for classification of CACS with threeclinical subtypes (Membranous, atrophic and hypertrophic), makes easier the study ofpatients with CACS and improves some aspects of Frieden's classification.

DE LA TESISSe estudian 28 casos de xantomatosis hiperlipoproteinemicas y normolipémicas mediante métodos clínicos, bioquímicos y cromatografía en capa fina del extracto lipídico de las lesiones cutáneas con resultados expresados en % merced al estudio densitométrico de las placas, así como en 36 casos de piel normal utilizada como patrón, además de estudios histológicos y ultraestructurales. Al mismo tiempo se hace una amplia revisión bibliográfica de las xantomatosis normolipémicas proponiendo una clasificación de las mismas según sus relaciones o no con enfermedades sistémicas acompañantes.

- / -

Little is known about the experiences of parental adjustment following the birth of a child with a facial birthmark due to a scarcity of literature. The literature within the fields of cleft lip and palate (CL/P) and cranio-facial abnormality (CFA) suggest that there are many factors involved in the process of parental adjustment. The aim of this project was to explore how parents adjust following the birth of their child with a facial birthmark. In-depth interviews were conducted with 13 parents of children aged between birth and two years old. The data was analysed using a Constructivist Grounded Theory approach and a model of adjustment was co-constructed from the data. It appears that parents move through a process of adjustment, that begins with coping with the initial shock and uncertainty before moving on to understanding the birthmark and treatment options, making sense of the birthmark, developing coping strategies and adjusting to birthmark. This process is affected by the availability of resources and the extent of social challenges experienced by the parents. The findings of this study indicate that psychology should be routinely involved with the assessment of parental psycho-social well-being and psycho-social support should be promoted for these families. Furthermore, greater training about birthmarks for medical professionals is recommended.

Loss-of-function mutations in the filaggrin gene(FLG) have recently been shown to be strongly associated with atopic dermatitis (AD). The overall aim of this study was to explore the role of filaggrin in skin barrier function and AD. There were two main focuses in this study. The first was a functional study whose primary objective was to determine if FLG mutations correlated with skin barrier dysfunction in AD. Fifty-five mild to moderate AD individuals were recruited, genotyped and had their skin barrier assessed using three different measures - transepidermal water loss (TEWL), skin capacitance and the number of tape strips required to abrogate skin barrier. A secondary aim of this functional study was to test the hypothesis that corneocytes were less adherent to one another in filaggrin-related AD compared to wild-type AD skin. The second main focus of this thesis was a structural study aimed at interrogating the structure-function relationship of filaggrin. Filaggrin protein was extracted and purified from a total of 21 AD and non-AD subjects and analysed using mass spectrometric techniques. Specifically, matrix assisted laser desorption/ionisation time-of-flight(MALDI-TOF)mass spectrometry (MS) and nano liquid chromatography tandem MS(LC-MS/MS) were utilised. Part of this structural study also involved developing and optimising the extraction and purification of filaggrin protein, including a novel way of extracting filaggrin from skin using tape stripping. In addition, novel filaggrin-specific enzyme-linked immunosorbant assay (ELISA) was also developed, which could serve as a useful screening test for the protein. In this study, FLG mutations were found to correlate with higher TEWL and fewer number of tape strips required to abrogate skin barrier, but not with skin capacitance. FLG mutations were also not shown to correlate with AD severity. The mean amount of protein extracted from filaggrin-related AD skin was also significantly higher compared to wild-type AD skin, supporting the hypothesis that corneocytes were less adherent to one another (and therefore, densely packed) in filaggrin-related AD skin. MS analysis of filaggrin confirmed the heterogeneic nature of filaggrin protein, even within a single individual. Interestingly, this structural study also showed that filaggrin was only minimally expressed in the skin of all the AD individuals studied, whether or not they possessed any FLG mutation. Due to the limited amount of filaggrin extracted from AD skin, it was not possible to conduct comparative structural analysis between filaggrin from AD and non-AD skin.