Participation of bcl -2 and bax in epidermal homeostasis and non-melanoma skin cancer

Non-melanoma skin cancer (NMSC) is the most frequently diagnosed form of cancer in United States. As in many other cancers, this slow growing malignancy manifests deregulated expression of apoptosis regulating proteins including bcl-2 family member proteins. To understand the role of apoptosis regulating protein in epidermal homeostasis and progression of NMSC, we investigated keratinocyte proliferation, differentiation and tumorigenesis in bcl-2 and bax null mice. The rate and the pattern of proliferation and spontaneous cell death were the same between the null and the control mice. Both bcl-2 and bax null epidermis showed decreased levels of cytokeratin 14 expression compared to the control littermates. Also, the gene knock out mice showed higher expression of cytokeratin 1 and loricrin in epidermis compared to the control mice. The apoptotic response to genotoxic agent, UV radiation (UVR), was assessed by counting sunburn cells. The bax null keratinocytes showed a resistance to apoptosis while bcl-2 null mice showed an increased susceptibility to cell death compared to the control mice. Moreover, we demonstrated an increase in tumor incidence in bax null mice compared to control littermates in the in vivo chemical carcinogenesis study. Next, we examined the tumor suppressor role of bax protein in NMSC by studying its participation in repair of UVR-mediated DNA lesions. In UVR treated primary keratinocytes from bax deficient mice, the level of CPD remaining was twice that of control cells at 48 hours. Similar results were obtained using embryonic fibroblasts from bax null and bax +/+ embryos, and also with a bax deficient prostate cancer cell line in which bax expression had been restored. However, the repair rate of 6-4 PP was unaffected by the absence of bax protein in all three of above mentioned cell types. In conclusion, bax protein may have a dual function in its role as tumor suppressor in NMSC. Bax may directly or indirectly facilitate DNA repair, or programmed cell death if DNA damage is too severe, thus, in either function, preserving genomic integrity following a genotoxic event. ^

Isolation and characterization of a novel human Mix -like homeobox gene MIXL

Molecular events involved in specification of early hematopoietic system are not well known. In Xenopus, a paired-box homeodomain family (Mix.1–4) has been implicated in this process. Although Mix-like homeobox genes have been isolated from zebrafish (bon), chicken (CMIX) and mice (MmI/MIXL1), isolation of a human Mix-like gene has remained elusive. ^ We have recently isolated and characterized a novel human Mix-like homeobox gene with a predicted open reading frame of 232 amino acids designated the Mix.1 homeobox (Xenopus laevis)-like gene (MIXL). The overall identity of this novel protein to CMIX and MmI/MIXL1 is 41% and 69%, respectively. However, the identity in the homeodomain is 66% to that of Xenopus Mix.1, 79% to that of CMIX, and 94% to that of MmI/MIXL1. In normal hematopoiesis, MIXL expression appears to be restricted immature B and T lymphoid cells. Several acute leukemic cell lines of B, T and myeloid lineages express MIXL suggesting a survival/block in differentiation advantage. Furthermore, Xenopus animal cap assay revealed that MIXL could induce expression of the α-globin gene, suggesting a functional conservation of the homeodomain. ^ Biochemical analysis revealed that MIXL proteins are phosphorylated at multiple sites. Immunoprecipitation and immunoblotting confirmed that MIXL is tyrosine phosphorylated. Mutational analysis determined that Tyr20 appears to be the site for phosphorylation. However, deletion analysis preliminarily showed that the proline-rich domain appears not to be necessary for tyrosine phosphorylation. The novel finding will help us make a deeper understanding of the regulation on homeodomain proteins by rarely reported tyrosine phosphorylation. ^ Taken together, isolation of the MIXL gene is the first step toward understanding novel regulatory circuits in early hematopoietic differentiation and malignant transformation. ^

Roles of p38 mitogen-activated protein kinase pathway in the regulation of SOX9 activity and chondrogenesis

Sox9 is a master transcription factor in chondrocyte differentiation. Several lines of evidence suggest that the p38 mitogen-activated protein kinase (MAPK) pathway is involved in chondrocyte differentiation. In the present study, we examined the roles of p38 in the regulation of SOX9 activity and chondrogenesis. ^ COS7 cells were transfected with a SOX9 expression vector and 4x48-p89, a luciferase construction harboring four tandem copies of a SOX9-dependent 48-bp enhancer in Col2a1. Coexpression of MKK6EE, a constitutively active mutant of MKK6, a MAPKK that specifically activates p38, further increased the activity of the SOX9-dependent 48-bp enhancer about 5-fold, and SOX9 protein levels were not increased under these conditions. This increase in enhancer activity was not observed in a mutant enhancer construct harboring mutations that abolish SOX9 binding. These data strongly suggested that activation of the p38 pathway results in increased activity of SOX9. In addition, the increase of the activity of the SOX9-dependent 48-bp enhancer by MKK6EE was also observed in primary chondrocytes, and this increase was abolished by coexpression of a p38 phosphatase, MKP5, and p38 specific inhibitors. Furthermore, treatment of primary chondrocytes with p38 inhibitors decreased the expression of Col2a1, a downstream target of Sox9, without affecting Sox9 RNA levels, further supporting the hypothesis that p38 plays a role in regulating Sox9 activity in chondrocytes. ^ To further study the role of the p38 MAPK pathway in chondrogenesis, we generated transgenic mice that express MKK6EE in chondrocytes under the control of the Col2a1 promoter/intron regulatory sequences. These mice showed a dwarf phenotype characterized by reduced chondrocyte proliferation and a delay in the formation of primary and secondary ossification centers. Histological analysis using in situ hybridization showed reduced expression of Indian hedgehog, PTH/PTHrP receptor, cyclin D1 and increased expression of p21. In addition, consistent with the notion that Sox9 activity was increased in these mice, transgenic mice that express MKK6EE in chondrocytes showed phenotypes similar to those of mice that overexpress SOX9 in chondrocytes. Therefore, our study provides in vivo evidence for the role of p38 in chondrocyte differentiation and suggests that Sox9 is a downstream target of the p38 MAPK pathway. ^

Nd and Sr isotopic composition of Cretaceous MORB

Chemical and isotopic (Nd and Sr) compositions have been determined for 12 Cretaceous basaltic samples (108 Ma old) from Holes 417D and 418A of Legs 51,52 and 53. We have found that: (1) The chemical compositions are typical of MORB. They do not vary systematically with the stratigraphic positions of the analyzed samples; thus, the chemical evolution is independent of the eruption sequence that occurred at this Cretaceous ridge. (2) REE patterns for all rocks are characterized by a strong LREE depletion with (La/Sm)N = 0.38-0.50; no significant Eu anomalies are found; HREE are nearly flat or slightly depleted towards Yb-Lu and have 12-18 * chondritic abundances. Combining the results of previous studies, it suggests that no significant temporal and spatial variation in magma chemistry (especially for LIL elements) has occurred in the 'normal' ridge segments over the last 150 Ma. (3) lsotopically, 143Nd/144Nd ratios vary from 0.513026 to 0.513154, corresponding to epsilon-Nd(0) = +7.5 to +10, and they fall in the typical range of MORB. However, these rocks have unexpectedly high 87Sr/86Sr ratios (0.70355-0.70470) which are attributed to the result of seawater-rock interaction. (4) The Nd model ages (Tin), ranging from 1.53 to 2.47 (average 2.06) AE, suggest that the upper mantle source(s) underwent a large scale chemical differentiation leading to LREE and other LIL element depletion about 2 AE ago, assuming a simple two-stage model. More realistically, the variation in Tm(Nd) or epsilon-Nd could be derived from mixing of heterogeneous mantle sources that were a consequence of continuous mantle differentiation and continental formation. (5) Because of the low mg values (0.52-0.63), the analyzed basaltic rocks do not represent primary liquids of mantle melting. The variation in La/Sm ratios and TiO2 are not compatible with a model in which all rocks are genetically related by a simple fractional crystallization. Rather, it is proposed that the basaltic rocks might have been derived from some heterogeneous upper mantle source with or without later magmatic mixing, and followed by some shallow-level fraetionations.

Territorial quality and sustainable development : Indicators of performance in the context of a model for participatory rural tourism certification

El turismo rural ha sido incorporado por pequeños establecimientos agropecuarios del partido de Cnel. Suárez, provincia de Buenos Aires, Argentina, como actividad alternativa para mejorar la calidad de vida de sus miembros y superar situaciones de crisis agravadas por la marginalidad productiva del SO bonaerense, región a la que pertenecen. Bajo el programa Cambio Rural del INTA (Instituto Nacional de Tecnología Agropecuaria) conforman el Grupo 'Cortaderas II', junto a otros emprendedores interesados en valorar el medio rural. Han avanzado en el proceso de reconocimiento de su identidad y puesta en valor de recursos específicos con anclaje en el territorio. Esta identidad comienza a apreciarse internamente, a raíz de la dinámica grupal lograda y la incipiente articulación con otros actores para la construcción de un partenariado público y privado que genere sinergias y contribuya al desarrollo sustentable del territorio. Sin embargo, aún no es claramente percibida por el turista, cada vez más exigente. Por lo tanto, el presente trabajo persigue proponer indicadores para evaluar el desempeño de un Sistema de Gestión de Calidad con enfoque territorial que, adaptando el modelo europeo 'Marca de Calidad Territorial', sustente una estrategia comercial de diferenciación del servicio y simultáneamente, mida el progreso hacia una mejor calidad de vida y fortalecimiento de vínculos con la cultura local y el entorno físico-natural en el marco del desarrollo sustentable. La investigación se plantea para la micro escala, ya que se trata de un estudio de caso, relevándose información primaria mediante observación directa y entrevistas semi-estructuradas, complementada con información secundaria diagnóstica utilizada por INTA. Las características del grupo y su dinámica de funcionamiento bajo el programa Cambio Rural revelan que es posible adoptar un proceso de certificación participativa propuesto para cuatro pilares de la calidad: de Bienes y Servicios, Institucional, Social y Ambiental. El modelo se integra con indicadores de evaluación de desempeño, agrupados en áreas clave para cada una de las dimensiones de la sustentabilidad, que contemplan el paisaje y la gestión de los recursos naturales; el impacto económico de la actividad, la calidad de la oferta y satisfacción del turista; así como las relaciones sociales internas y los vínculos con otros actores del territorio. Principalmente se encontraron fortalezas en la búsqueda de partenariados y debilidades en aspectos de comunicación y promoción. Se considera que este sistema de herramientas de gestión sustentable permitiría superar las dificultades de una certificación individual, pudiendo aplicarse a emprendimientos con otra ubicación geográfica

Territorial quality and sustainable development : Indicators of performance in the context of a model for participatory rural tourism certification

El turismo rural ha sido incorporado por pequeños establecimientos agropecuarios del partido de Cnel. Suárez, provincia de Buenos Aires, Argentina, como actividad alternativa para mejorar la calidad de vida de sus miembros y superar situaciones de crisis agravadas por la marginalidad productiva del SO bonaerense, región a la que pertenecen. Bajo el programa Cambio Rural del INTA (Instituto Nacional de Tecnología Agropecuaria) conforman el Grupo 'Cortaderas II', junto a otros emprendedores interesados en valorar el medio rural. Han avanzado en el proceso de reconocimiento de su identidad y puesta en valor de recursos específicos con anclaje en el territorio. Esta identidad comienza a apreciarse internamente, a raíz de la dinámica grupal lograda y la incipiente articulación con otros actores para la construcción de un partenariado público y privado que genere sinergias y contribuya al desarrollo sustentable del territorio. Sin embargo, aún no es claramente percibida por el turista, cada vez más exigente. Por lo tanto, el presente trabajo persigue proponer indicadores para evaluar el desempeño de un Sistema de Gestión de Calidad con enfoque territorial que, adaptando el modelo europeo 'Marca de Calidad Territorial', sustente una estrategia comercial de diferenciación del servicio y simultáneamente, mida el progreso hacia una mejor calidad de vida y fortalecimiento de vínculos con la cultura local y el entorno físico-natural en el marco del desarrollo sustentable. La investigación se plantea para la micro escala, ya que se trata de un estudio de caso, relevándose información primaria mediante observación directa y entrevistas semi-estructuradas, complementada con información secundaria diagnóstica utilizada por INTA. Las características del grupo y su dinámica de funcionamiento bajo el programa Cambio Rural revelan que es posible adoptar un proceso de certificación participativa propuesto para cuatro pilares de la calidad: de Bienes y Servicios, Institucional, Social y Ambiental. El modelo se integra con indicadores de evaluación de desempeño, agrupados en áreas clave para cada una de las dimensiones de la sustentabilidad, que contemplan el paisaje y la gestión de los recursos naturales; el impacto económico de la actividad, la calidad de la oferta y satisfacción del turista; así como las relaciones sociales internas y los vínculos con otros actores del territorio. Principalmente se encontraron fortalezas en la búsqueda de partenariados y debilidades en aspectos de comunicación y promoción. Se considera que este sistema de herramientas de gestión sustentable permitiría superar las dificultades de una certificación individual, pudiendo aplicarse a emprendimientos con otra ubicación geográfica

Territorial quality and sustainable development : Indicators of performance in the context of a model for participatory rural tourism certification

El turismo rural ha sido incorporado por pequeños establecimientos agropecuarios del partido de Cnel. Suárez, provincia de Buenos Aires, Argentina, como actividad alternativa para mejorar la calidad de vida de sus miembros y superar situaciones de crisis agravadas por la marginalidad productiva del SO bonaerense, región a la que pertenecen. Bajo el programa Cambio Rural del INTA (Instituto Nacional de Tecnología Agropecuaria) conforman el Grupo 'Cortaderas II', junto a otros emprendedores interesados en valorar el medio rural. Han avanzado en el proceso de reconocimiento de su identidad y puesta en valor de recursos específicos con anclaje en el territorio. Esta identidad comienza a apreciarse internamente, a raíz de la dinámica grupal lograda y la incipiente articulación con otros actores para la construcción de un partenariado público y privado que genere sinergias y contribuya al desarrollo sustentable del territorio. Sin embargo, aún no es claramente percibida por el turista, cada vez más exigente. Por lo tanto, el presente trabajo persigue proponer indicadores para evaluar el desempeño de un Sistema de Gestión de Calidad con enfoque territorial que, adaptando el modelo europeo 'Marca de Calidad Territorial', sustente una estrategia comercial de diferenciación del servicio y simultáneamente, mida el progreso hacia una mejor calidad de vida y fortalecimiento de vínculos con la cultura local y el entorno físico-natural en el marco del desarrollo sustentable. La investigación se plantea para la micro escala, ya que se trata de un estudio de caso, relevándose información primaria mediante observación directa y entrevistas semi-estructuradas, complementada con información secundaria diagnóstica utilizada por INTA. Las características del grupo y su dinámica de funcionamiento bajo el programa Cambio Rural revelan que es posible adoptar un proceso de certificación participativa propuesto para cuatro pilares de la calidad: de Bienes y Servicios, Institucional, Social y Ambiental. El modelo se integra con indicadores de evaluación de desempeño, agrupados en áreas clave para cada una de las dimensiones de la sustentabilidad, que contemplan el paisaje y la gestión de los recursos naturales; el impacto económico de la actividad, la calidad de la oferta y satisfacción del turista; así como las relaciones sociales internas y los vínculos con otros actores del territorio. Principalmente se encontraron fortalezas en la búsqueda de partenariados y debilidades en aspectos de comunicación y promoción. Se considera que este sistema de herramientas de gestión sustentable permitiría superar las dificultades de una certificación individual, pudiendo aplicarse a emprendimientos con otra ubicación geográfica

Chemical composition, age, and magnetic parameters of igneous rocks from the Franz Josef Land

A combined study of magnetic parameters of basalt and andesite samples has been carried out in the framework of geological investigations of the Franz Josef Land. This study has included determination of coercivity, saturation magnetization, Curie points, natural remanent magnetization (NRM), and magnetic susceptibility as well as examination of ferromagnetic minerals with a microscope. Data on chemical composition of the rocks have been obtained for all the samples, and radiological ages have been determined for the majority of the rocks. Thermomagnetic curves of the samples have been subdivided into four types depending on composition of ferromagnetic NRM carriers. Data showing multiple changes in the predominant composition of the igneous rocks have been obtained. Each stage of magmatism is characterized by a specific type of the ferromagnetic component in the rocks and, therefore, magnetomineralogical investigations can be used for differentiation and correlation of the igneous rocks.

Uranium and radioactive isotopes in bottom sediments and Fe-Mn nodules and crusts of seas and oceans

The main stages of the sedimentary cycle of uranium in modern marine basins are under consideration in the book. Annually about 18 thousand tons of dissolved and suspended uranium enters the ocean with river runoff. Depending on a type of a marine basin uranium accumulated either in sediments of deep-sea basins, or in sediments of continental shelves and slopes. In the surface layer of marine sediments hydrogenic uranium is predominantly bound with organic matter, and in ocean sediments also with iron, manganese and phosphorus. In diagenetic processes there occurs partial redistribution of uranium in sediments, as well as its concentration in iron-manganese, phosphate and carbonate nodules and biogenic phosphate detritus. Concentration of uranium in marine sediments of various types depending on their composition, as well as on forms of its entering, degree of differentiation and of sedimentation rates, on hydrochemical regime and water circulation, and on intensity of diagenetic processes.

Termination and cost analysis: Complexity and precision issues

The research in this thesis is related to static cost and termination analysis. Cost analysis aims at estimating the amount of resources that a given program consumes during the execution, and termination analysis aims at proving that the execution of a given program will eventually terminate. These analyses are strongly related, indeed cost analysis techniques heavily rely on techniques developed for termination analysis. Precision, scalability, and applicability are essential in static analysis in general. Precision is related to the quality of the inferred results, scalability to the size of programs that can be analyzed, and applicability to the class of programs that can be handled by the analysis (independently from precision and scalability issues). This thesis addresses these aspects in the context of cost and termination analysis, from both practical and theoretical perspectives. For cost analysis, we concentrate on the problem of solving cost relations (a form of recurrence relations) into closed-form upper and lower bounds, which is the heart of most modern cost analyzers, and also where most of the precision and applicability limitations can be found. We develop tools, and their underlying theoretical foundations, for solving cost relations that overcome the limitations of existing approaches, and demonstrate superiority in both precision and applicability. A unique feature of our techniques is the ability to smoothly handle both lower and upper bounds, by reversing the corresponding notions in the underlying theory. For termination analysis, we study the hardness of the problem of deciding termination for a speci�c form of simple loops that arise in the context of cost analysis. This study gives a better understanding of the (theoretical) limits of scalability and applicability for both termination and cost analysis.

Connexin-43 in the osteogenic BM niche regulates its cellular composition and the bidirectional traffic of hematopoietic stem cells and progenitors

Connexin-43 (Cx43), a gap junction protein involved in control of cell proliferation, differentiation and migration, has been suggested to have a role in hematopoiesis. Cx43 is highly expressed in osteoblasts and osteogenic progenitors (OB/P). To elucidate the biologic function of Cx43 in the hematopoietic microenvironment (HM) and its influence in hematopoietic stem cell (HSC) activity, we studied the hematopoietic function in an in vivo model of constitutive deficiency of Cx43 in OB/P. The deficiency of Cx43 in OB/P cells does not impair the steady state hematopoiesis, but disrupts the directional trafficking of HSC/progenitors (Ps) between the bone marrow (BM) and peripheral blood (PB). OB/P Cx43 is a crucial positive regulator of transstromal migration and homing of both HSCs and progenitors in an irradiated microenvironment. However, OB/P Cx43 deficiency in nonmyeloablated animals does not result in a homing defect but induces increased endosteal lodging and decreased mobilization of HSC/Ps associated with proliferation and expansion of Cxcl12-secreting mesenchymal/osteolineage cells in the BM HM in vivo. Cx43 controls the cellular content of the BM osteogenic microenvironment and is required for homing of HSC/Ps in myeloablated animals

Detection and Tracking of Dynamic Objects. A MultiRobot approach to Critical Infrastructures Surveillance.

Despite that Critical Infrastructures (CIs) security and surveillance are a growing concern for many countries and companies, Multi Robot Systems (MRSs) have not been yet broadly used in this type of facilities. This dissertation presents a novel study of the challenges arisen by the implementation of this type of systems and proposes solutions to specific problems. First, a comprehensive analysis of different types of CIs has been carried out, emphasizing the influence of the different characteristics of the facilities in the design of a security and surveillance MRS. One of the most important needs for the surveillance of a CI is the detection of intruders. From a technical point of view this problem can be abstracted as equivalent to the Detection and Tracking of Mobile Objects (DATMO). This dissertation proposes algorithms to solve this specific problem in a CI environment. Using 3D range images of the environment as input data, two detection algorithms for ground robots have been developed. These detection algorithms provide a list of moving objects in the robot detection area. Direct image differentiation and computer vision techniques are used when the robot is static. Alternatively, multi-layer ground reconstructions are compared to detect the dynamic objects when the robot is moving. Since CIs usually spread over large areas, it is very useful to incorporate aerial vehicles in the surveillance MRS. Therefore, a moving object detection algorithm for aerial vehicles has been also developed. This algorithm compares the real optical flow obtained from a down-face oriented camera with an artificial optical flow computed using a RANSAC based homography matrix. Two tracking algorithms have been developed to follow the moving objects trajectories. These algorithms can efficiently handle occlusions and crossings, as well as exchange information among robots. The multirobot tracking can be applied to any type of communication structure: centralized, decentralized or a combination of both. Even more, the developed tracking algorithms are independent of the detection algorithms and could be potentially used with other detection procedures or even with static sensors, such as cameras. In addition, using the 3D point clouds available to the robots, a relative localization algorithm has been developed to improve the position estimation of a given robot with observations from other robots. All the developed algorithms have been extensively tested in different simulated CIs using the Webots robotics simulator. Furthermore, the algorithms have also been validated with real robots operating in real scenarios. In conclusion, this dissertation presents a multirobot approach to Critical Infrastructure Surveillance, mainly focusing on Detecting and Tracking Dynamic Objects.

Sparse differential resultant formulas: between the linear and the nonlinear case

A matrix representation of the sparse differential resultant is the basis for efficient computation algorithms, whose study promises a great contribution to the development and applicability of differential elimination techniques. It is shown how sparse linear differential resultant formulas provide bounds for the order of derivation, even in the nonlinear case, and they also provide (in many cases) the bridge with results in the nonlinear algebraic case.

Development in Rural Areas Through Capacity Building and Education for Business

This paper is based on a case study located in Avila, central Spain. Its main objectives are to implement an entrepreneurship program and design a plan of capacity building and education for business in order to promote the development of rural areas. The methodological approach of the program is based on the use of tools that permit involving the various actors of the area from the early planning stages. The university's group that is carrying out the field work has relied on these participatory tools in very different areas and contexts for over 25 years. This has allowed the development of an advanced planning model called ?Working With People? that connects expert and experience knowledge in the territories where it is applied. With this methodological approach, the diagnosis of the territory and the design of the program's strategy has been carried out. Once completed the first phase of the program and in order to ensure the sustainability and applicability of future entrepreneurial initiatives, it is necessary to support and strengthen potential entrepreneurs through training activities and capacity building. It relies on ?How to learn from people who live there? to promote investment projects and to teach them with adequate educational skills. In this context, this article aims to study the implementation strategy of these training and capacity building activities studied from an academic perspective, as well as analyzing the potential effects of these actions in promoting entrepreneurship in the territories

Regulation of the human p21/WAF1/Cip1 promoter in hepatic cells by functional interactions between Sp1 and Smad family members

The cell cycle inhibitor p21/WAF1/Cip1 is expressed in many cell types and is regulated by p53-dependent and p53-independent mechanisms. p21 is an important regulator of hepatocyte cell cycle, differentiation, and liver development, but little is known about the regulation of its synthesis in hepatocytes. We report herein that the p21 gene is constitutively expressed in human hepatoma HepG2 cells. Deletion analysis of the p21 promoter showed that it contains a distal (positions −2,300/−210) and a proximal (positions −124 to −61) region that act synergistically to achieve high levels of constitutive expression. The proximal region that consists of multiple Sp1 binding sites is essential for constitutive p21 promoter activity in hepatocytes. This region also mediates the transcriptional activation of the p21 promoter by members of the Smad family of proteins, which play important role in the transduction of extracellular signals such as transforming growth factor β, activin, etc. Constitutive expression of p21 was severely reduced by a C-terminally truncated form of Smad4 that was shown previously to block signaling through Smads. Smad3/4 and to a much lesser extent Smad2/4 caused high levels of transcriptional activation of the p21 promoter. Transactivation was compromised by N- or C-terminally truncated forms of Smad3. By using Gal4-Sp1 fusion proteins, we show that Smad proteins can activate gene transcription via functional interactions with the ubiquitous factor Sp1. These data demonstrate that Smad proteins and Sp1 participate in the constitutive or inducible expression of the p21 gene in hepatic cells.