Regulation of human (adrenal) androgen biosynthesis-New insights from novel throughput technology studies

Androgens are precursors for sex steroids and are predominantly produced in the human gonads and the adrenal cortex. They are important for intrauterine and postnatal sexual development and human reproduction. Although human androgen biosynthesis has been extensively studied in the past, exact mechanisms underlying the regulation of androgen production in health and disease remain vague. Here, the knowledge on human androgen biosynthesis and regulation is reviewed with a special focus on human adrenal androgen production and the hyperandrogenic disorder of polycystic ovary syndrome (PCOS). Since human androgen regulation is highly specific without a good animal model, most studies are performed on patients harboring inborn errors of androgen biosynthesis, on human biomaterials and human (tumor) cell models. In the past, most studies used a candidate gene approach while newer studies use high throughput technologies to identify novel regulators of androgen biosynthesis. Using genome wide association studies on cohorts of patients, novel PCOS candidate genes have been recently described. Variant 2 of the DENND1A gene was found overexpressed in PCOS theca cells and confirmed to enhance androgen production. Transcriptome profiling of dissected adrenal zones established a role for BMP4 in androgen synthesis. Similarly, transcriptome analysis of human adrenal NCI-H295 cells identified novel regulators of androgen production. Kinase p38α (MAPK14) was found to phosphorylate CYP17 for enhanced 17,20 lyase activity and RARB and ANGPTL1 were detected in novel networks regulating androgens. The discovery of novel players for androgen biosynthesis is of clinical significance as it provides targets for diagnostic and therapeutic use.

Calidad del 'sitio' en el oasis norte, Mendoza (Argentina)

La disminución del agua destinada al uso agrícola, la salinización de los acuíferos subterráneos y el advenimiento de la tecnología de Sistemas de Información Geográfica (SIG) han posibilitado conocer la calidad de los sitios, fundamentalmente los riesgos de salinización de los suelos del oasis del río Mendoza-Argentina. El presente trabajo se fundamenta en dos estudios anteriores: uno de relevantamiento de suelos y el otro de análisis de calidad de aguas subterráneas. En el primero se efectúo la actualización del relevantamiento de suelos del río Mendoza usando SIG. El muestreo de suelos y los análisis físicos (textura) y químicos (salinidad, conductividad eléctrica) se realizaron en 1974. Los lugares de muestreo y sus atributos, graficados como cobertura de puntos, se extrapolaron a sus zonas de influencia convirtiéndolos en polígonos y posteriormente se rasterizaron. El segundo trabajo fue la digitalización y georreferenciación, también al sistema de coordenadas Universal Transverse Mercator (UTM), de los mapas de las curvas de isosalinidad. La salinidad está medida por la conductividad eléctrica específica del agua subterránea de los tres niveles de explotación que existen en la cuenca norte de Mendoza. El monitoreo se realizó en el período 1990/1991. Las isolíneas, posteriormente, fueron rasterizadas. Con los procesos de superposición y tabulación cruzada de los SIG se integraron las diversas "capas" de datos de suelos y calidades de aguas subterráneas y se generaron mapas temáticos que expresan la clasificación y localización regional de calidades del sitio, basado fundamentalmente en los riesgos de salinización de los suelos.

(Table 2) Concentrations of dissolved Zn, Cd, Pb, and Cu in waters of the Baltic Sea

A method of determination of low concentrations of zinc, cadmium, lead, and copper dissolved in seawater and interstitial water with use of inversion voltammetry with AC solution conditions is described. The optimum conditions for measurements with accumulation on a mercury-film electrode with a silver substrate are presented. Detection limit is 0.2 ?g/l for zinc and 0.05 ?g/l for cadmium, lead, and copper. Large number of determinations can be carried out during expeditions at natural pH of seawater containing these four metals.

Dive depth and dive duration data of female southern elephant seals from Marion Island between 2004 and 2008 with links to datasets

The at-sea behaviour of marine top predators provides valuable insights into the distribution of prey species and strategies used by predators to exploit patchily distributed resources. We describe the water column usage and dive strategies of female southern elephant seals from Marion Island tracked between 2004 and 2008. Dives representing increases in forage effort were identified using a method that combines dive type analyses and the calculation of relative amounts of time that animals spend in the bottom phases of dives. Results from this analysis indicate that female elephant seals from Marion Island tend to display lower levels of forage effort closer to the island and display intensive opportunistic forage bouts that occur at a minimum distance of approximately 215 km from the island. Females from Marion Island dived deeper and for longer periods of time, compared to females from other populations. Most animals displayed positive diel vertical migration, evidently foraging pelagically on vertically migrating prey. A few animals displayed periods of reverse (negative) diel vertical migration, however, diving to deeper depths at night, compared to daytime. This behaviour is difficult to explain and prey species targeted during such periods unknown. Our results illustrate plasticity in foraging behaviour of southern elephant seals, as well as inter-population differences in forage strategies.

Metodología para la gestión integral de las presas de laminación de avenidas y su aplicación a la cuenca hidrográfica del Segura

En el levante español se producen periódicamente intensos aguaceros (gota fría), que generan grandes riadas, desbordándose los cauces y asolando los campos y las ciudades, ocasionando graves daños en la economía local y con frecuencia, por desgracia, víctimas personales. La Confederación Hidrográfica del Segura ha puesto en marcha un vasto sistema de protección, conocido como Plan de Defensa contra Avenidas, del que ya se encuentran en explotación un conjunto de presas cuyo objetivo prioritario es la laminación de las avenidas. Las presas de laminación ubicadas en esta área mediterránea son estructuras muy singulares, al estar ubicadas en cuencas extremadamente secas, con alta pendiente, rápida respuesta, picos altos de caudal, y gran poder erosivo del flujo. Las aguas arrastran en suspensión un alto porcentaje de aporte sólido y flotantes, que quedan retenidos en el embalse, aterrándolo e inutilizando los desagües. En esta tesis doctoral se analizan los criterios de actuación para la gestión de estas presas de laminación, con objeto de que se puedan introducir en las Normas de Explotación innovaciones destinadas a mejorar la eficiencia del conjunto, de manera que se consiga la máxima protección posible y al mismo tiempo se puedan aprovechar las aguas, sin menoscabo de su objetivo prioritario, para otros fines complementarios. Como consecuencia del trabajo realizado se indica la conveniencia de establecer un conjunto de normas de actuación comunes a todas las presas del sistema, como es la estrategia de recibir siempre a los hidrogramas de la riada con las compuertas de los desagües de fondo cerrados, o la conveniencia de que en todas las presas se dispongan compuertas en estos órganos. También se dictan recomendaciones en la gestión del vaciado, para conseguir el máximo aprovechamiento del agua, introduciendo el concepto innovador de los trasvases virtuales. Asimismo, se analiza la conveniencia de fijar normas diferenciales específicas atendiendo a las peculiaridades de cada presa, en cuanto se refiere al nivel máximo de explotación, al cumplimiento de los condicionantes ambientales, al mantenimiento de la calidad del agua, y a otros aspectos singulares que caracterizan a cada uno de estos embalses. The Spanish Mediterranean region hydrology is characterised by low rainfall, but concentrated in extreme convective events. Those heavy rains and storms lead to large, flash-floods which cause devastation of fields, villages and, unfortunately, loss of lives. Segura River Basin Authority has developed an ambitious plan for flood protection and control that is constituted by a system of flood protection dams, river channelling, river diversions diversions and levees. Flood protection dams placed on this area, are distinct structures, as they are located on extremely dry basins, with high slopes and limited vegetal covering. These facts, along with the very high rainfall intensity, are determinants of a rapid response and a very erosive flow. The flow transports a large rate of solids in suspension, and also debris, which many times block and clog the outlets intakes, disabling them. In such a case, the dam would be inoperative, in an emergency state and remedial measures should be taken. This PhD thesis analyses the flood protection dams operation criteria, for providing managers a methodology to analyse each individual case and also the flood protection system as a whole. Additionally practical rules and recommendations, to cope with the objectives of dam safety, flood protection and water resources use, are given. The rules consider the specific differences and risks that may be faced by the dams. To that end, they were grouped, depending on their characteristics: allowance for permanent water storage, environmental restrictions, water quality, geotechnical problems, type of structure, etc. As a result of this research, it is recommended to design these dams with valves for regulating the bottom outlets. The availability of valves is linked with the routing strategy of receiving flood with the outlets closed. This has been shown as the best measure to prevent clogging and opens the possibility of storing temporarily the water, making then a scheduled emptying of the reservoir, which could be computed as a virtual transfer to the regulating dams. Emptying should be done in accordance with the situation of the other dams of the system and of the rivers downstream. This detention time would make possible also the beginning of the sedimentation process which is necessary for using the water with the modern irrigation systems that are in use in this area.

Mechanism-based inhibition of the melatonin rhythm enzyme: Pharmacologic exploitation of active site functional plasticity

Serotonin N-acetyltransferase is the enzyme responsible for the diurnal rhythm of melatonin production in the pineal gland of animals and humans. Inhibitors of this enzyme active in cell culture have not been reported previously. The compound N-bromoacetyltryptamine was shown to be a potent inhibitor of this enzyme in vitro and in a pineal cell culture assay (IC50 ≈ 500 nM). The mechanism of inhibition is suggested to involve a serotonin N-acetyltransferase-catalyzed alkylation reaction between N-bromoacetyltryptamine and reduced CoA, resulting in the production of a tight-binding bisubstrate analog inhibitor. This alkyltransferase activity is apparently catalyzed at a functionally distinct site compared with the acetyltransferase activity active site on serotonin N-acetyltransferase. Such active site plasticity is suggested to result from a subtle conformational alteration in the protein. This plasticity allows for an unusual form of mechanism-based inhibition with multiple turnovers, resulting in “molecular fratricide.” N-bromoacetyltryptamine should serve as a useful tool for dissecting the role of melatonin in circadian rhythm as well as a potential lead compound for therapeutic use in mood and sleep disorders.

Structure of the HIV-1 integrase catalytic domain complexed with an inhibitor: A platform for antiviral drug design

HIV integrase, the enzyme that inserts the viral DNA into the host chromosome, has no mammalian counterpart, making it an attractive target for antiviral drug design. As one of the three enzymes produced by HIV, it can be expected that inhibitors of this enzyme will complement the therapeutic use of HIV protease and reverse transcriptase inhibitors. We have determined the structure of a complex of the HIV-1 integrase core domain with a novel inhibitor, 5ClTEP, 1-(5-chloroindol-3-yl)-3-hydroxy-3-(2H-tetrazol-5-yl)-propenone, to 2.1-Å resolution. The inhibitor binds centrally in the active site of the integrase and makes a number of close contacts with the protein. Only minor changes in the protein accompany inhibitor binding. This inhibitor complex will provide a platform for structure-based design of an additional class of inhibitors for antiviral therapy.

Rescue of abasic hammerhead ribozymes by exogenous addition of specific bases.

We have synthesized 13 hammerhead ribozyme variants, each containing an abasic residue at a specific position of the catalytic core. The activity of each of the variants is significantly reduced. In four cases, however, activity can be rescued by exogenous addition of the missing base. For one variant, the rescue is 300-fold; for another, the rescue is to the wild-type level. This latter abasic variant (G10.1X) has been characterized in detail. Activation is specific for guanine, the base initially removed. In addition, the specificity for guanine versus adenine is substantially altered by replacing C with U in the opposite strand of the ribozyme. These results show that a binding site for a small, noncharged ligand can be created in a preexisting ribozyme structure. This has implications for structure-function analysis of RNA, and leads to speculations about evolution in an "RNA world" and about the potential therapeutic use of ribozymes.

Oligodeoxynucleotides inhibit retinal neovascularization in a murine model of proliferative retinopathy.

Diseases characterized by retinal neovascularization are among the principal causes of visual loss worldwide. The hypoxia-stimulated expression of vascular endothelial growth factor (VEGF) has been implicated in the proliferation of new blood vessels. We have investigated the use of antisense phosphorothioate oligodeoxynucleotides against murine VEGF to inhibit retinal neovascularization and VEGF synthesis in a murine model of proliferative retinopathy. Intravitreal injections of two different antisense phosphorothioate oligodeoxynucleotides prior to the onset of proliferative retinopathy reduced new blood vessel growth a mean of 25 and 31% compared with controls. This inhibition was dependent on the concentration of antisense phosphorothioate oligodeoxynucleotides and resulted in a 40-66% reduction in the level of VEGF protein, as determined by Western blot analysis. Control (sense, nonspecific) phosphorothioate oligodeoxynucleotides did not cause a significant reduction in retinal neovascularization or VEGF protein levels. These data further establish a fundamental role for VEGF expression in ischemia-induced proliferative retinopathies and a potential therapeutic use for antisense phosphorothioate oligodeoxynucleotides.

El futuro de la regulación del sector del abastecimiento en España

La gestión del servicio de abastecimiento de agua es un tema de muy relevante y de plena actualidad, no solo en España sino a nivel mundial. En España, desde la forma de prestación del servicio a través de la gestión directa o la gestión indirecta, o el tipo en sí de gestión indirecta que se desarrolle –empresa privada, pública o mixta- dan como resultado prácticamente tantas situaciones como ayuntamientos existen ya que la competencia es municipal, según reconoce la Ley de Bases de Régimen Local. La heterogeneidad de circunstancias de gestión de los recursos hídricos, la fragmentación de un sector clave y la falta de indicadores comparables entre diferentes opciones para la prestación de este servicio, parece indicar la necesidad de apostar por la creación de un ente regulador independiente dada la naturaleza monopolística del sector del abastecimiento y el saneamiento, junto con el carácter esencial del bien que se gestiona. Mientras que las instituciones gubernamentales dictan las políticas generales a través de leyes y reglamentos, los operadores -públicos, privados o mixtos- son los responsables de prestar los servicios a la población, y sería el órgano regulador quien velaría por el cumplimiento de las normas. El MAGRAMA está estudiando la posibilidad de elaborar una Ley sobre el Ciclo Integral del Agua de Uso Urbano con las dificultades que esto tiene en consideración a las competencias de las Comunidades Autónomas, lo que sin duda ayudaría a homologar la gestión del agua urbana en España, regulando un monopolio natural, ahora en manos de más de 8.100 municipios y un reducido número de empresas concesionarias, auténticas detentadoras de estos servicios. En este trabajo tratamos de analizar las ventajas que supondría para España contar con una agencia de regulación similar a las existentes en numerosos países, tanto europeos como de América Latina.

Commonplace book of Moses Appleton, 1791-1815 (inclusive)

Volume containing medicinal recipes, medical notes, poetry, and obituaries written by Dr. Moses Appleton (1773-1849). Many of the recipes were copied from medical texts or other publications. His "cure for the dropsy," taken from the New York Herald, contained stale cider, parsley, horseradish, oxymel squills (sea onion in honey), and juniper berries. For diarrhea, he prescribed a blackberry syrup. Several entries indicate Appleton practiced Thomsonian medicine, an alternative system based on use of botanicals. The medical notes include an account of his treatment of a man with smallpox in 1815, and entries on patients he inoculated with cowpox matter. Another entry dated in 1796 provides instructions from the Massachusetts Humane Society for "treatment to be used with persons apparently dead from drowning," which included blowing tobacco smoke in the victim's lungs and applying warm blankets for several hours. Appleton adds a note questioning whether or not the lungs also should be "often artificially inflated." There is additionally a history of prominent physicians dating from ancient Greece.

Cases copied by James Lloyd from Mr. Steed, apothecary in Guy's Hospital, London, 1751-1787 (inclusive)

Contains medical cases copied by James Lloyd (1728-1810), primarily between 1751 and 1754, from Mr. Steed, an apothecary at Guy's Hospital in London, England. The volume has additional medical cases dating from 1780 to 1787. Lloyd transcribed the names, ages, and symptoms of the patients, as well as the medicines and medical care delivered to them. The volume is divided into chapters based on the type of case, which included vision loss; fluor albus, or leucorrhoea; diabetes; and dysentery. There is also a letter pasted into the volume addressed to Dr. Brigham of the Boston Medical Library Association from Lloyd's great-grandson, dated 4 November 1887.

Worcester, Massachusetts, ca. 1860 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of the city of Worcester, Mass. : from actual surveys under the direction of P. Ball, C.E. It was published by Smith & Mc.Kinney, between 1850 and 1860. Scale [ca. 1:3,600]. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Massachusetts State Plane Coordinate System, Mainland Zone (in Feet) (Fipszone 2001) coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, public buildings, schools, industry locations (e.g. mills, factories, etc.), selected private buildings with names of property owners, town and ward boundaries, cemeteries, and more. Includes also engravings of important buildings and advertisements in margins. This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Brookfield, North Brookfield, West Brookfield, & East Brookfield, Massachusetts, 1855 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of the towns of Brookfield, North Brookfield, West Brookfield, Worcester County, Mass, surveyed & drawn by L. Fagan. It was published by Richard Clark in 1855. Scale [ca. 1:21,120]. Covers the towns of Brookfield, East Brookfield, North Brookfield, and West Brookfield, Massachusetts. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Massachusetts State Plane Coordinate System, Mainland Zone (in Feet) (Fipszone 2001) coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads, drainage, public buildings, schools, churches, cemeteries, industry locations (e.g. mills, factories, mines, etc.), selected private buildings with names of property owners, town boundaries, and more. Relief shown by hachures. Includes also town center insets and selected building illustrations.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.