Homeostatic transcriptional regulation of bcl-2 in the epidermis

Bcl-2, a crucial regulator of cell survival, is frequently overexpressed in basal cell carcinomas (BCCs), the most commonly diagnosed cancers. Regulation of bcl-2 expression in epidermal keratinocytes is not well characterized. In the epidermis, bcl-2 is expressed only in keratinocytes of the basal layer and the outer root sheath of hair follicles and no bcl-2 expression in suprabasalar keratinocytes. The calcium gradient in the epidermis is a potent regulator of keratinocyte differentiation. Increasing calcium concentrations associated with differentiation, resulted in the downregulation of a 2.9 kb bcl-2 promoter luciferase construct. The AP-1 family of transcription factors is differentially expressed in the strata of the epidermis and has been shown to be involved in the stage specific expression of numerous differentiation markers in the epidermis. In silico analysis of the bcl-2 promoter and gene reporter assays showed that co-transfection of JUNB and JUND, but not other AP-1 dimers, caused a significant upregulation of the bcl-2 promoter in primary keratinocytes. Immunoelectrophoretic mobility shift assays, in vivo chromatin immunoprecipitation (ChIP) studies and mutational analysis of AP-1 binding site 3 on the bcl-2 promoter identified it as the site involved in bcl-2 regulation. Utilizing site directed mutants, we determined that phosphorylation at Ser90/Ser100 residues of JUND is required for the activation of the bcl-2 promoter. ^ The sonic hedgehog (SHH) pathway is frequently deregulated in BCCs and, we have shown that GLI1 upregulates bcl-2 in keratinocytes. While examining potential regulation of the SHH pathway extracellular calcium, we found that higher calcium concentrations are associated with lowered HH pathway activity and upregulation of suppressor of fused (SUFU) which negatively regulates the SHH pathway. ChIP assays, and in vivo mouse models, show that ΔNp63α, a crucial regulator of epidermal development, binds and activates the SUFU promoter in differentiating keratinocytes. Increasing SUFU levels prevent transactivation of the bcl-2 promoter. In vitro SUFU knockdown along with in vivo SUFU+/− murine models demonstrate a significant upregulation of bcl-2 expression. ^ In conclusion, the spatial and temporal expression of bcl-2 during keratinocyte differentiation in the epidermis is a complex process requiring cooperative interactions of specific signaling cascades and transcription factors. ^

Genetic epidemiology of childhood brain tumors

A rare familial cancer syndrome involving childhood brain tumors (CBT), breast cancer, sarcomas and an array of other tumors has been described (Li and Fraumeni 1969, 1975, 1982, 1987). A survey of CBT identified through the Connnecticut Tumor Registry in 1984 revealed a high frequency of CBT, leukemia and other childhood cancer in siblings of CBT patients (Farwell and Flannery, 1984). Other syndromes such as neurofibromatosis and nevoid basal cell carcinoma syndrome have also been associated with CBT; however, no systematic family studies have been conducted to determine the extent to which cancer aggregates in family members of CBT patients. This family study was designed to determine the frequency of cancer aggregation overall or at specific sites, to determine the frequency of known or potentially hereditary syndromes in families of CBT patients, and to determine a genetic model to characterize familial cancer syndromes and to identify specific kindreds to which such a model(s) might apply. This study includes 244 confirmed CBT patients referred to the University of Texas M. D. Anderson Cancer Center between the years 1944 and 1983, diagnosed under the age of 15 years and resident in the U.S. or Canada. Family histories were obtained on the proband's first (parents, siblings and offspring) and second degree (proband's aunts, uncles and grandparents) relatives following sequential sampling scheme rules. To determine if cancer aggregates in families, we compared the cancer experience in the population to that expected in the general population using Connecticut Tumor Registry calendar year, age, race and sex-specific rates. The standardized incidence ratio (SIR) for cancer overall was 0.91 (41 observed (O) and 44.94 expected (E); 95% Confidence Interval (CI) = 0.65-1.24). We observed a significant excess of colon cancer among the proband's first degree relatives (O/E = 5/1.64; 95% CI = 1.01-7.65), in particular those under age 45 year. Segregation analysis showed evidence for multifactorial inheritance in the small percentage (N = 5) of the families. ^

Contribución al estudio de propiedades ópticas en medios heterogéneos

Como contribución del estudio de medios heterogéneos, esta tesis recoge el trabajo llevado a cabo sobre modelado teórico y simulación del estudio de las propiedades ópticas de la piel y del agua del mar, como ejemplos paradigmáticos de medios heterogéneos. Se ha tomado como punto de partida el estudio de la propagación de la radiación óptica, más concretamente de la radiación láser, en un tejido biológico. La importancia de la caracterización óptica de un tejido es fundamental para manejar la interacción radiación-tejido que permite tanto el diagnóstico como la terapéutica de enfermedades y/o de disfunciones en las Ciencias de la Salud. Sin olvidar el objetivo de ofrecer una metodología de estudio, con un «enfoque ingenieril», de las propiedades ópticas en un medio heterogéneo, que no tiene por qué ser exclusivamente el tejido biológico. Como consecuencia de lo anterior y de la importancia que tiene el agua dentro de los tejidos biológicos se decide estudiar en otro capítulo las propiedades ópticas del agua dentro de un entorno heterogéneo como es el agua del mar. La selección del agua del mar, como objeto de estudio adicional, es motivada, principalmente, porque se trata de un sistema heterogéneo fácilmente descriptible en cada uno de sus elementos y permite evaluar una amplia bibliografía. Además se considera que los avances que han tenido lugar en los últimos años en las tecnologías fotónicas van a permitir su uso en los métodos experimentales de análisis de las aguas. El conocimiento de sus propiedades ópticas permite caracterizar los diferentes tipos de aguas de acuerdo con sus compuestos, así como poder identificar su presencia. Todo ello abre un amplio abanico de aplicaciones. En esta tesis doctoral, se ha conseguido de manera general: • Realizar un estudio del estado del arte del conocimiento de las propiedades ópticas de la piel y la identificación de sus elementos dispersores de la luz. • Establecer una metodología de estudio que nos permita obtener datos sobre posibles efectos de la radiación en los tejidos biológicos. •Usar distintas herramientas informáticas para simular el transporte de la radiación laser en tejidos biológicos. • Realizar experimentos mediante simulación de láser, tejidos biológicos y detectores. • Comparar los resultados conocidos experimentalmente con los simulados. • Estudiar los instrumentos de medida de la respuesta a la propagación de radiación laser en tejidos anisotrópicos. • Obtener resultados originales para el diagnóstico y tratamiento de pieles, considerando diferente razas y como alteración posible en la piel, se ha estudiado la presencia del basalioma. • Aplicación de la metodología de estudio realizada en la piel a la simulación de agua de mar. • Obtener resultados originales de simulación y análisis de cantidad de fitoplancton en agua; con el objetivo de facilitar la caracterización de diferentes tipos de aguas. La tesis doctoral se articula en 6 capítulos y 3 anexos perfectamente diferenciados con su propia bibliografía en cada uno de ellos. El primer capítulo está centrado en la problemática del difícil estudio y caracterización de los medios heterogéneos debidos a su comportamiento no homogéneo y anisotrópico ante las radiaciones ópticas. Así pues, presentaremos una breve introducción al comportamiento tanto de los tejidos como del océano ante radiaciones ópticas y definiremos sus principales propiedades: la absorción, el scattering, la anisotropía y los coeficientes de reflexión. Como continuación, un segundo capítulo trata de acercarnos a la resolución del problema de cómo caracterizar las propiedades ópticas descritas en el primer capítulo. Para ello, primero se introducen los modelos teóricos, en segundo lugar los métodos de simulación más empleados y, por último, enumerar las principales técnicas de medida de la propagación de la luz en los tejidos vivos. El tercer capítulo, centrado en la piel y sus propiedades, intenta realizar una síntesis de lo que se conoce sobre el comportamiento de la piel frente a la propagación de las radiaciones ópticas. Se estudian sus elementos constituyentes y los distintos tipos de pieles. Por último se describe un ejemplo de aplicación más inmediata que se beneficia de este conocimiento. Sabemos que el porcentaje de agua en el cuerpo humano es muy elevado, en concreto en la piel se considera de aproximadamente un 70%. Es obvio, por tanto, que conocer cómo afecta el agua en la propagación de una radiación óptica facilitaría el disponer de patrones de referencia; para ello, se realiza el estudio del agua del mar. En el cuarto capítulo se estudian las propiedades del agua del mar como medio heterogéneo de partículas. En este capítulo presentamos una síntesis de los elementos más significativos de dispersores en el océano, un estudio de su comportamiento individual frente a radiaciones ópticas y su contribución al océano en su conjunto. Finalmente, en el quinto capítulo se describen los resultados obtenidos en los distintos tipos de simulaciones realizadas. Las herramientas de simulación empleadas han sido las mismas tanto para el caso del estudio de la piel como para el agua del mar, por ello ambos resultados son expuestos en el mismo capítulo. En el primer caso se analizan diferentes tipos de agua oceánica, mediante la variación de las concentraciones de fitoplancton. El método empleado permite comprobar las diferencias que pueden encontrarse en la caracterización y diagnóstico de aguas. El segundo caso analizado es el de la piel; donde se estudia el comportamiento de distintos tipos de piel, se analizan para validar el método y se comprueba cómo el resultado es compatible con aplicaciones, actualmente comerciales, como la de la depilación con láser. Como resultado significativo se muestra la posible metodología a aplicar para el diagnóstico del cáncer de piel conocido como basalioma. Finalmente presentamos un capítulo dedicado a los trabajos futuros basados en experimentación real y el coste asociado que implicaría el llevarlo a cabo. Los anexos que concluyen la tesis doctoral versan por un lado sobre el funcionamiento del vector común de toda la tesis: el láser, sus aplicaciones y su control en la seguridad y por otro presentamos los coeficientes de absorción y scattering que hemos utilizado en nuestras simulaciones. El primero condensa las principales características de una radiación láser desde el punto de vista de su generación, el segundo presenta la seguridad en su uso y el tercero son tablas propias, cuyos parámetros son los utilizados en el apartado de experimentación. Aunque por el tipo de tesis que defiendo no se ajusta a los modelos canónicos de tesis doctoral, el lector podrá encontrar en esta tesis de forma imbricada, el modelo común a todas las tesis o proyectos de investigación con una sección dedicada al estado del arte con ejemplos pedagógicos para facilitar la compresión y se plantean unos objetivos (capítulos 1-4), y un capítulo que se subdivide en materiales y métodos y resultados y discusiones (capítulo 5 con sus subsecciones), para finalizar con una vista al futuro y los trabajos futuros que se desprenden de la tesis (capítulo 6). ABSTRACT As contribution to the study of heterogeneous media, this thesis covers the work carried out on theoretical modelling and simulation study of the optical properties of the skin and seawater, as paradigmatic examples of heterogeneous media. It is taken as a starting point the study of the propagation of optical radiation, in particular laser radiation in a biological tissue. The importance of optical characterization of a tissue is critical for managing the interaction between radiation and tissues that allows both diagnosis and therapy of diseases and / or dysfunctions in Health Sciences. Without forgetting the aim of providing a methodology of study, with "engineering approach" of the optical properties in a heterogeneous environment, which does not have to be exclusively biological tissue. As a result of this and the importance of water in biological tissues, we have decided to study the optical properties of water in a heterogeneous environment such as seawater in another chapter. The selection of sea water as an object of further study is motivated mainly because it is considered that the advances that have taken place in recent years in photonic technologies will allow its use in experimental methods of water analysis. Knowledge of the optical properties to characterize the different types of waters according to their compounds, as well as to identify its presence. All of this opens a wide range of applications. In this thesis, it has been generally achieved: • Conduct a study of the state of the art knowledge of the optical properties of the skin and identifying its light scattering elements. • Establish a study methodology that allows us to obtain data on possible effects of radiation on biological tissues. • Use different computer tools to simulate the transport of laser radiation in biological tissues. • Conduct experiments by simulating: laser, detectors, and biological tissues. • Compare the known results with our experimentally simulation. • Study the measuring instruments and its response to the propagation of laser radiation in anisotropic tissues. • Get innovative results for diagnosis and treatment of skin, considering different races and a possible alteration in the skin that we studied: the presence of basal cell carcinoma. • Application of the methodology of the study conducted in the skin to simulate seawater. • Get innovative results of simulation and analysis of amount of phytoplankton in water; in order to facilitate the characterization of different types of water. The dissertation is divided into six chapters and three annexes clearly distinguished by their own literature in each of them. The first chapter is focused on the problem of difficult study and characterization of heterogeneous media due to their inhomogeneous and anisotropic behaviour of optical radiation. So we present a brief introduction to the behaviour of both tissues at the cellular level as the ocean, to optical radiation and define the main optical properties: absorption, scattering, anisotropy and reflection coefficients. Following from this, a second chapter is an approach to solving the problem of how to characterize the optical properties described in the first chapter. For this, first the theoretical models are introduced, secondly simulation methods more used and, finally, the main techniques for measuring the propagation of light in living tissue. The third chapter is focused on the skin and its properties, tries to make a synthesis of what is known about the behaviour of the skin and its constituents tackle the spread of optical radiation. Different skin types are studied and an example of immediate application of this knowledge benefits described. We know that the percentage of water in the human body is very high, particularly in the skin is considered about 70%. It is obvious, therefore, that knowing how the water is affected by the propagation of an optical radiation facilitate to get reference patterns; For this, the study of seawater is performed. In the fourth chapter the properties of seawater as a heterogeneous component particles are studied. This chapter presents a summary of the scattering elements in the ocean, its individual response to optical radiation and its contribution to the ocean as a whole. In the fifth chapter the results of the different types of simulations are described. Simulation tools used were the same for the study of skin and seawater, so both results are presented in the chapter. In the first case different types of ocean water is analysed by varying the concentrations of phytoplankton. The method allows to check the differences that can be found in the characterization and diagnosis of water. The second case analysed is the skin; where the behaviour of different skin types are studied and checked how the result is compatible with applications currently trade, such as laser hair removal. As a significant result of the possible methodology to be applied for the diagnosis of skin cancer known as basal cell carcinoma is shown. Finally we present a chapter on future work based on actual experimentation and the associated cost which it would involve carrying out. The annexes conclude the thesis deal with one hand on the functioning of the common vector of the whole thesis: laser, control applications and safety and secondly we present the absorption and scattering coefficients we used in our simulations. The first condenses the main characteristics of laser radiation from the point of view of their generation, the second presents the safety in use and the third are own tables, whose parameters are used in the experimental section. Although the kind of view which I advocate does not meet the standard models doctoral thesis, the reader will find in this thesis so interwoven, the common model to all theses or research projects with a section on the state of the art pedagogical examples to facilitate the understanding and objectives (Chapters 1-4), and a chapter is divided into materials and methods and results and discussions (Chapter 5 subsections) arise, finishing with a view to the future and work future arising from the thesis (Chapter 6).

Suspensor-derived polyembryony caused by altered expression of valyl-tRNA synthetase in the twn2 mutant of Arabidopsis

The twn2 mutant of Arabidopsis exhibits a defect in early embryogenesis where, following one or two divisions of the zygote, the decendents of the apical cell arrest. The basal cells that normally give rise to the suspensor proliferate abnormally, giving rise to multiple embryos. A high proportion of the seeds fail to develop viable embryos, and those that do, contain a high proportion of partially or completely duplicated embryos. The adult plants are smaller and less vigorous than the wild type and have a severely stunted root. The twn2-1 mutation, which is the only known allele, was caused by a T-DNA insertion in the 5′ untranslated region of a putative valyl-tRNA synthetase gene, valRS. The insertion causes reduced transcription of the valRS gene in reproductive tissues and developing seeds but increased expression in leaves. Analysis of transcript initiation sites and the expression of promoter–reporter fusions in transgenic plants indicated that enhancer elements inside the first two introns interact with the border of the T-DNA to cause the altered pattern of expression of the valRS gene in the twn2 mutant. The phenotypic consequences of this unique mutation are interpreted in the context of a model, suggested by Vernon and Meinke [Vernon, D. M. & Meinke, D. W. (1994) Dev. Biol. 165, 566–573], in which the apical cell and its decendents normally suppress the embryogenic potential of the basal cell and its decendents during early embryo development.

Characterization of two patched receptors for the vertebrate hedgehog protein family

The multitransmembrane protein Patched (PTCH) is the receptor for Sonic Hedgehog (Shh), a secreted molecule implicated in the formation of embryonic structures and in tumorigenesis. Current models suggest that binding of Shh to PTCH prevents the normal inhibition of the seven-transmembrane-protein Smoothened (SMO) by PTCH. According to this model, the inhibition of SMO signaling is relieved after mutational inactivation of PTCH in the basal cell nevus syndrome. Recently, PTCH2, a molecule with sequence homology to PTCH, has been identified. To characterize both PTCH molecules with respect to the various Hedgehog proteins, we have isolated the human PTCH2 gene. Biochemical analysis of PTCH and PTCH2 shows that they both bind to all hedgehog family members with similar affinity and that they can form a complex with SMO. However, the expression patterns of PTCH and PTCH2 do not fully overlap. While PTCH is expressed throughout the mouse embryo, PTCH2 is found at high levels in the skin and in spermatocytes. Because Desert Hedgehog (Dhh) is expressed specifically in the testis and is required for germ cell development, it is likely that PTCH2 mediates its activity in vivo. Chromosomal localization of PTCH2 places it on chromosome 1p33–34, a region deleted in some germ cell tumors, raising the possibility that PTCH2 may be a tumor suppressor in Dhh target cells.

Disruption of the FG nucleoporin NUP98 causes selective changes in nuclear pore complex stoichiometry and function

The NUP98 gene encodes precursor proteins that generate two nucleoplasmically oriented nucleoporins, NUP98 and NUP96. By using gene targeting, we have selectively disrupted the murine NUP98 protein, leaving intact the expression and localization of NUP96. We show that NUP98 is essential for mouse gastrulation, a developmental stage that is associated with rapid cell proliferation, but dispensable for basal cell growth. NUP98−/− cells had an intact nuclear envelope with a normal number of embedded nuclear pore complexes. Typically, NUP98-deficient cells contained on average approximately 5-fold more cytoplasmic annulate lamellae than control cells. We found that a set of cytoplasmically oriented nucleoporins, including NUP358, NUP214, NUP88, and p62, assembled inefficiently into nuclear pores of NUP98−/− cells. Instead, these nucleoporins were prominently associated with the annulate lamellae. By contrast, a group of nucleoplasmically oriented nucleoporins, including NUP153, NUP50, NUP96, and NUP93, had no affinity for annulate lamellae and assembled normally into nuclear pores. Mutant pores were significantly impaired in transport receptor-mediated docking of proteins with a nuclear localization signal or M9 import signal and showed weak nuclear import of such substrates. In contrast, the ability of mutant pores to import ribosomal protein L23a and spliceosome protein U1A appeared intact. These observations show that NUP98 disruption selectively impairs discrete protein import pathways and support the idea that transport of distinct import complexes through the nuclear pore complex is mediated by specific subsets of nucleoporins.

Complete Cytolysis and Neonatal Lethality in Keratin 5 Knockout Mice Reveal Its Fundamental Role in Skin Integrity and in Epidermolysis Bullosa Simplex

In human patients, a wide range of mutations in keratin (K) 5 or K14 lead to the blistering skin disorder epidermolysis bullosa simplex. Given that K14 deficiency does not lead to the ablation of a basal cell cytoskeleton because of a compensatory role of K15, we have investigated the requirement for the keratin cytoskeleton in basal cells by inactivating the K5 gene in mice. We report that the K5−/− mice die shortly after birth, lack keratin filaments in the basal epidermis, and are more severely affected than K14−/− mice. In contrast to the K14−/− mice, we detected a strong induction of the wound-healing keratin K6 in the suprabasal epidermis of cytolyzed areas of postnatal K5−/− mice. In addition, K5 and K14 mice differed with respect to tongue lesions. Moreover, we show that in the absence of K5 and other type II keratins, residual K14 and K15 aggregated along hemidesmosomes, demonstrating that individual keratins without a partner are stable in vivo. Our data indicate that K5 may be the natural partner of K15 and K17. We suggest that K5 null mutations may be lethal in human epidermolysis bullosa simplex patients.

Expression of cyclin D1 in epithelial tissues of transgenic mice results in epidermal hyperproliferation and severe thymic hyperplasia.

To study the involvement of cyclin D1 in epithelial growth and differentiation and its putative role as an oncogene in skin, transgenic mice were developed carrying the human cyclin D1 gene driven by a bovine keratin 5 promoter. As expected, all squamous epithelia including skin, oral mucosa, trachea, vaginal epithelium, and the epithelial compartment of the thymus expressed aberrant levels of cyclin D1. The rate of epidermal proliferation increased dramatically in transgenic mice, which also showed basal cell hyperplasia. However, epidermal differentiation was unaffected, as shown by normal growth arrest of newborn primary keratinocytes in response to high extracellular calcium. Moreover, an unexpected phenotype was observed in the thymus. Transgenic mice developed a severe thymic hyperplasia that caused premature death due to cardio-respiratory failure within 4 months of age. By 14 weeks, the thymi of transgenic mice increased in weight up to 40-fold, representing 10% of total body weight. The hyperplastic thymi had normal histology revealing a well-differentiated cortex and medulla, which supported an apparently normal T-cell developmental program based on the distribution of thymocyte subsets. These results suggest that proliferation and differentiation of epithelial cells are under independent genetic controls in these organs and that cyclin D1 can modulate epithelial proliferation without altering the initiation of differentiation programs. No spontaneous development of epithelial tumors or thymic lymphomas was perceived in transgenic mice during their first 8 months of life, although they continue under observation. This model provides in vivo evidence of the action of cyclin D1 as a pure mediator of proliferation in epithelial cells.

Overexpression of sonic hedgehog suppresses embryonic hair follicle morphogenesis

The Sonic Hedgehog (Shh) signalling pathway plays a central role in the development of the skin and hair follicle and is a major determinant of skin tumorigenesis, most notably of basal cell carcinoma (BCC). Various mouse models involving either ablation or overexpression of key members of the Shh signalling pathway display a range of skin tumours. To further examine the role of Shh in skin development. we have overexpressed Shh in a subset of interfollicular basal cells from 12.5 dpc under the control of the human keratin 1 (HK1) promoter. The HK1-Shh transgenic mice display a range of skin anomalies, including highly pigmented inguinal lesions and regions of alopecia. The most striking hair follicle phenotype is a suppression in embryonic follicle development between 14.0 and 19.0 dpc, resulting in a complete absence of guard, awl, and auchene hair fibres. These data indicate that alternative signals are responsible for the development of different hair follicles and point to a major role of Shh signalling in the morphogenesis of guard, awl, and auchene hair fibres. Through a comparison with other mouse models, the characteristics of the HK1-Shh transgenic mice suggest that the precise timing and site of Shh expression are key in dictating the resultant skin and tumour phenotype. 2003 Elsevier Inc. All rights reserved.

Equine laminitis: glucose deprivation and MMP activation induce dermo-epidermal separation in vitro

Reasons for performing study: Acute laminitis is characterised by hoof lamellar dermal-epidermal separation at the basement membrane (BM) zone. Hoof lamellar explants cultured in vitro can also be made to separate at the basement membrane zone and investigating how this occurs may give insight into the poorly understood pathophysiology of laminitis. Objectives: To investigate why glucose deprivation and metalloproteinase (MMP) activation in cultured lamellar explants leads to dermo-epidermal separation. Methods: Explants, cultured without glucose or with the MMP activator p-amino-phenol-mercuric acetate (APMA), were subjected to tension and processed for transmission electron microscopy (TEM). Results: Without glucose, or with APMA, explants under tension separated at the dermo-epidermal junction. This in vitro separation occurred via 2 different ultrastructural processes. Lack of glucose reduced hemidesmosomes (HDs) numbers until they disappeared and the basal cell cytoskeleton collapsed. Anchoring filaments (AFs), connecting the basal cell plasmalemma to the BM, were unaffected although they failed under tension. APMA activation of constituent lamellar MMPs did not affect HDs but caused AFs to disappear, also leading to dermo-epidermal separation under tension. Conclusions: Natural laminitis may occur in situations where glucose uptake by lamellar basal cells is compromised (e.g. equine Cushing's disease, obesity, hyperlipaemia, ischaemia and septicaemia) or when lamellar MMPs are activated (alimentary carbohydrate overload). Potential relevance: Therapies designed to facilitate peripheral glucose uptake and inhibit lamellar MMP activation may prevent or ameliorate laminitis.

Polymorphisms in glutathione S-transferases and non-melanoma skin cancer risk in Australian renal transplant recipients

Caucasian renal transplant recipients from Queensland, Australia have the highest non-melanoma skin cancer (NMSC) risk worldwide. Although ultraviolet light (UVR) exposure is critical, genetic factors also appear important. We and others have shown that polymorphism in the glutathione S-transferases (GST) is associated with NMSC in UK recipients. However, the effect of high UVR exposure and differences in immunosuppressive regimen on these associations is unknown. In this study, we examined allelism in GSTM1, GSTM3, GSTT1 and GSTP1 in 361 Queensland renal transplant recipients. Data on squamous (SCC) and basal cell carcinoma (BCC), UVR/tobacco exposure and genotype were obtained. Associations with both NMSC risk and numbers were examined using logistic and negative binomial regression, respectively. In the total group, GSTM1 AB [P = 0.049, rate ratio (RR) = 0.23] and GSTM3 AA (P = 0.015, RR = 0.50) were associated with fewer SCC. Recipients were then stratified by prednisolone dose (less than or equal to7 versus >7 mg/day). In the low-dose group, GSTT1 null (P = 0.006, RR = 0.20) and GSTP1 Val/Val (P = 0.021, RR = 0.20) were associated with SCC numbers. In contrast, in the high-dose group, GSTM1 AB (P = 0.009, RR = 0.05), GSTM3 AB (P = 0.042, RR = 2.29) and BB (P = 0.014, RR = 5.31) and GSTP1 Val/Val (P = 0.036, RR = 2.98) were associated with SCC numbers. GSTM1 AB (P = 0.016) and GSTP1 Val/Val (P = 0.046) were also associated with fewer BCC in this group. GSTP1 associations were strongest in recipients with lower UVR/tobacco exposure. The data confirm our UK findings, suggesting that protection against UVR-induced oxidative stress is important in NMSC development in recipients, but that this effect depends on the immunosuppressant regimen.

Histopathology of oligofructose-induced acute laminitis in heifers

Histopathology of the dermo-epidermal junction in the lamellar region of front claws was examined in 6 dairy heifers given an alimentary oligofructose overload and compared with sections from a control group of 6 heifers. Four of the 6 heifers administered oligofructose developed clinical signs of acute laminitis before they were euthanized. Postmortem samples from front claws were processed for histology. Eleven histopathologic characteristics were selected from the existing literature and used in a blinded evaluation of sections. In total, 104 front claw samples, including 8 samples from 2 cows having spontaneously occurring acute laminitis, were evaluated histologically using hematoxylin and eosin as well as periodic acid-Schiff staining. The major morphological features associated with oligofructose-induced acute clinical laminitis were stretching of lamellae, dermal edema, hemorrhage, changes in basal cell morphology, presence of white blood cells in dermis, and signs of basement membrane detachment. Changes at the lamellar junction of claw tissue affected by oligofructose-induced clinical laminitis resembled tissue from the 2 cows suffering from spontaneous acute clinical laminitis, and generally were consistent with existing descriptions of laminitis histopathology. Important exceptions to existing descriptions in the literature were stretching of lamellae and basement membrane changes. Not previously described, we considered these early signs of acute laminitis. In conclusion, this study documents that oligofructose-induced clinical laminitis is associated with histopathological changes at the lamellar interface. A weakened dermo-epidermal junction is a possible intermediate stage in the pathophysiology of bovine sole ulceration at the typical site.

Clinical outcomes from skin screening clinics within a community-based melanoma screening program

Background : Within a randomized trial of population screening for melanoma, primary care physicians conducted whole-body skin examinations and referred all patients with suspect lesions to their own doctor for further treatment. Objective: Our aim was to describe characteristics of skin screening participants, clinical screening diagnoses, management following referral, and specificity and yield of screening examinations. Methods: Information collected from consent forms, referral forms, and histopathological reports of lesions that had been excised or undergone biopsy was analyzed by means of descriptive statistics. Results: A total of 16,383 whole-body skin examinations resulted in 2302 referrals (14.1% overall; 15.5% men, 18.2% >= 50 years of age) for 4129 suspect lesions (including 222 suspected melanoma, 1101 suspected basal cell carcinomas [BCCs], 265 suspected squamous cell carcinomas [SCCs]). Histopathologic results were available for 94.8% of 1417 lesions excised and confirmed 33 melanomas (23 in men; 24 in participants ? 50 years of age), 259 BCCs, and 97 SCCs. The probability of detecting skin cancer of any type within the program was 2.4%. The estimated specificity of whole-body skin examinations for melanoma was 86.1% (95% confidence interval = 85.6-86.6). The positive predictive value (number of confirmed/number of lesions excised or biopsied x 100) for melanoma was 2.5%, 19.3% for BCC, and 7.2% for SCC (overall positive predictive value for skin cancer, 28.9%). Limitations: Follow-up of participants with a negative screening examination has not been conducted for the present investigation. Conclusions: The rate of skin cancer detected per 100 patients screened was higher than previously reported and men and attendees older than 50 years more frequently received a referral and diagnosis of melanoma. The specificity for detection of melanoma through whole-body skin examination by a primary care physician was comparable to that of other screening tests, including mammography.

Long-term increase in sunscreen use in an Australian community after a skin cancer prevention trial

Background. Given the public health burden of skin cancer in white populations, an increase in sun protective behavior is needed. In a highrisk community, we assessed long-term Sunscreen use among people who had participated in a randomized trial of daily Sunscreen application for prevention of skin cancer. Methods. In 1992, 1621 residents of the subtropical Australian township of Nambour were randomly allocated to either daily or discretionary sunscreen use until 1996. From 1997 to 2002, we monitored by questionnaires their ongoing sunscreen use. Results. People who had never or irregularly used sunscreen when in summer sun before the trial were more likely (P < 0.0001) to be sustaining regular application especially to their face (20% vs. 11%) and forearms (14% vs. 5%) if they had been allocated to daily, not discretionary, use of sunscreen for 5 years. Conclusions. Regular voluntary sunscreen use for skin cancer prevention can be sustained by sun-sensitive people in the long term. Habit formation appears to be an important goal for sun protection programs among those living, or on vacation, in sunny places. (c) 2005 Elsevier Inc. All rights reserved.