Two Highly Conserved Cysteine Residues in HPV16 L2 Form an Intramolecular Disulfide Bond and Are Critical for Infectivity in Human Keratinocytes

Minor capsid protein L2 performs an indispensable but uncharacterized role in human papillomavirus infections. A neutralizing B cell epitope has recently been mapped to the N-terminus of HPV16 L2, residues 17-36, and exposure of this region of L2 has been implicated in translocation of incoming virions from the endo/lysosomal compartment to the cellular cytoplasm. Here we examine the redox state of Cys22 and Cys28 two highly conserved cysteines located within this epitope. We also investigate the infectivity of virions containing L2 single and double cysteine point mutants.Denaturing/non-reducing gel analysis and thiol labeling experiments of wild type and cysteine mutant HPV16 virion particles strongly support the existence of a buried intramolecular C22-C28 disulfide bond. The disulfide was confirmed by tandem mass spectrometry of L2 protein from non-reduced virions. Single C22S and C28S and the double C22/28S mutants were non-infectious but had no apparent defects in cell binding, endocytosis, or trafficking to lysosomes by 8 h post infection. During infection with L2 mutant particles, there was a marked decrease in L2 levels compared to wild type L2-containing virions, suggesting a failure of mutant L2/genome complexes to exit the endo/lysosomal compartment.L2 residues C22 and C28 are bound as an intramolecular disulfide bond in HPV16 virions and are necessary for infectivity. Previous work has suggested that the furin-dependent exposure of the 17-36 epitope and subsequent interaction of this region with an unknown receptor is necessary for egress from the endo/lysosomal compartment and infection. Identification of the C22-C28 disulfide suggests that reduction of this disufide bond may be necessary for exposure of 17-36 and HPV16 infection

Fokus-Naturtag - Beratungskonzept zur Förderung des Naturschutzbewusstseins und der Naturschutzleistungen landwirtschaftlicher Betriebe

Das Beratungskonzept Fokus-Naturtag wurde während einer 3-jährigen Projektlaufzeit in Kooperation mit staatlichen und nicht-staatlichen Einrichtungen aus den Bereichen Landwirtschaft und Naturschutz in drei Bundesländern entwickelt, anhand von 100 landwirtschaftlichen Testbetrieben mit ökologischer und konventioneller Betriebsweise erprobt und ausgewertet. Ziel des Vorhabens war es, Landwirte als Gestalter der Kulturlandschaft durch eine komprimierte Beratung für Naturschutzbelange zu sensibilisieren, neue Denkanstöße zu setzen und ihre Motivation für Naturschutzbelange zu steigern. Mit dem Fokus-Naturtag steht nun erstmals ein Beratungsbaukasten mit speziell aufeinander abgestimmten Komponenten zur Verfügung, der es ermöglicht, eine Naturschutz- oder Biodiversitätsberatung auf landwirtschaftlichen Betrieben innerhalb eines überschaubaren zeitlichen und finanziellen Rahmens anzubieten. Das Baukastenprinzip ermöglicht dabei die flexible Praxisanwendung der einzelnen Bestandteile, wenn notwendig oder gewünscht auch unabhängig voneinander. Der entwickelte Beratungsansatz ist kompatibel mit einer von der EU geforderten und finanziell geförderten Beratung im Bereich Umwelt- und Naturschutz. Die im letzten Projektjahr durchgeführte Evaluierung diente einer ex-post-Bewertung des Fokus-Naturtags sowie des Beratungskonzepts Kulturlandplan. Die Evaluierung hatte zum Ziel, die Wirksamkeit der Beratungen hinsichtlich der Umsetzung vorgeschlagener Naturschutzmaßnahmen und der Sensibilisierung für Naturschutzbelange zu ermitteln. Darüber hinaus wurde die Methodik des Beratungskonzepts bewertet und Faktoren, die eine Umsetzung der vorgeschlagenen Maßnahmen auf betrieblicher Ebene begünstigen oder behindern, identifiziert

O MOVIMENTO SEM TERRA E O LETRAMENTO NO MEIO RURAL BRASILEIRO

In this article we discuss the conceptions of literacy produced by MST (Landless Movement) and its influence over one rural public school teacher. We analyse her literacies practices, recognizing that her political involvement and professional formation up dates her practices, observing the interactional processes built in his classroom and the ways she deals with reading and writting in schooling processes. We conclude that the practices observed in the researched classroom offer us opportunity to reafirm the necessity of more dialogical school practices and community involvement in local school project

Bone Marrow Mesenchymal Stem Cells Provide an Antibiotic-Protective Niche for Persistent Viable Mycobacterium tuberculosis that Survive Antibiotic Treatment

During tuberculosis (TB), some Mycobacterium tuberculosis bacilli persist in the presence of an active immunity and antibiotics that are used to treat the disease. Herein, by using the Cornell model of TB persistence, we further explored our recent finding that suggested that M. tuberculosis can escape therapy by residing in the bone marrow (BM) mesenchymal stem cells. We initially showed that M. tuberculosis rapidly disseminates to the mouse BM after aerosol exposure and maintained a stable burden for at least 220 days. In contrast, in the lungs, the M. tuberculosis burden peaked at 28 days and subsequently declined approximately 10-fold. More important, treatment of the mice with the antibiotics rifampicin and isoniazid, as expected, resulted in effective clearance of M. tuberculosis from the lungs and spleen. In contrast, M. tuberculosis persisted, albeit at low numbers, in the BM of antibiotic-treated mice. Moreover, most viable M. tuberculosis was recovered from the bone marrow CD271+CD45−-enriched cell fraction, and only few viable bacteria could be isolated from the CD271−CD45+ cell fraction. These results clearly show that BM mesenchymal stem cells provide an antibiotic-protective niche for M. tuberculosis and suggest that unraveling the mechanisms underlying this phenomenon will enhance our understanding of M. tuberculosis persistence in treated TB patients

Imagem e História no Cemitério São Miguel da Cidade de Goiás

Resumo: este trabalho apresenta algumas reflexões referentes às histórias reproduzidas dentro do cemitério da Cidade de Goiás. Alguns aspectos formais são percebíveis nas diversificações das imagens que foram produzidas ao longo do tempo, a partir daí, analiso alguns conceitos que ajudam na relevância das imagens e de seus significados que ficam entre a história e o imaginário. A história da â??menina da xícaraâ? é apresentada para mostrar como imagem e imaginário transforma a história local trazendo novos sentidos. Palavras-chave: Imagem. Representações. Cemitérios. Imaginári

Propuesta de implementación de un sistema MRP para reducir los costos de inventario de materia prima en la producción de alimentos balanceados para pollos en Molino El Cortijo S.A.C.

El presente trabajo tuvo como objetivo general la Propuesta de Implementación de un Sistema MRP para reducir los costos de inventario de materia prima en la Producción de Alimento Balanceado para Pollos en Molino El Cortijo S.A.C. Con este propósito se siguieron pasos definidos entre los cuales se hallaban el analizar el estado actual de la gestión de inventarios en la organización, determinar el nivel de ventas y el cumplimiento de la producción, examinar los niveles de inventario actuales y determinar los costos de inventario relacionados con la producción de alimento balanceado para pollos. Luego era propuesta la metodología de planificación de materiales MRP, con el fin de aprovechar la oportunidad de mejora, la cual se relaciona a la reducción de los costos de inventario. Los resultados que se lograron fueron, reducir el costo de mantener inventarios de materia prima de S/. 271,661.10 Nuevos Soles en el año 2013 o S/. 22,638.43 Nuevos Soles al mes a S/. 207,367.14 al año o S/. 17,280.59 Nuevos Soles al mes (un aproximado de 23.67% de ahorro). Se redujo el costo de emisión de orden de compra de S/.108.41 Nuevos Soles en el año 2013 a S/.105.74 Nuevos Soles, esto debido al aumento de órdenes de compra emitidas. Se redujo asimismo el costo total de inventario, de S/. 605,859.72 en el año 2013 a S/. 416,478.55 Nuevos Soles según la propuesta, generándose un ahorro total de 31.26%. Este ahorro involucra el costo de ordenar los pedidos de aprovisionamiento y el costo anual de mantener inventario por materia prima y producto terminado. De igual manera, los indicadores económicos para la implementación de la presente propuesta exponen los siguientes resultados: VAN (Valor actual neto) de S/.12 933.55; TIR (Tasa interna de retorno) por 119.99%; PRI (Periodo de recuperación de la inversión) de 2.0 años y un indicador de B/C (Beneficio - costo) por 2.1.ABSTRACT The present work has the overall aim of Proposed Implementation of a MRP system to reduce inventory costs of raw material in the production of Balanced Feed for Chickens in Molino El Cortijo SAC. For this purpose I defined steps which were to analyze the current state of inventory management in the organization, determine the level of sales and fulfillment production, review current inventory levels and inventory costing followed related to the production of balanced feed for chickens. Then it was proposed methodology MRP (materials planning), in order to seize the opportunity for improvement, which is related to reduced inventory costs. The results achieved were, reduce the cost of maintaining inventories of raw materials from S /. 271,661.10 Nuevos Soles in 2013 or S/. 22,638.43 soles a month to S /. 207,367.14 a year or S/. 17,280.59 nuevos soles per month (an approximately 23.67% savings). The cost of issuing purchase order S/ .108.41 Nuevos Soles in 2013 to S / .105.74 Nuevos Soles declined, this due to increased purchase orders issued. The total cost of inventory, was also reduced of S/. 605,859.72 in 2013 to S /. 416,478.55 Nuevos Soles as proposed, generating a total savings of 31.26%. This involves saving the cost of issuing purchase orders and the annual cost of maintaining inventory raw material and finished product. Similarly, economic indicators for the implementation of this proposal presented the following results: NPV (net present value) of S/ .12, 933.55; IRR (Internal Rate of Return) by 119.99%; PRI (Period of return on investment) of 2.0 years and a indicator of B/C (Benefit – Cost) for 2.1

Papel de MTS1 y S5H en la respuesta defensiva de plantas de tomate frente a la infección con la bacteria Pseudomonas syringae

Throughout evolution, plants have been developing defence systems against different abiotic and biotic aggressions by their environment. These defensive systems include both constitutive and inducible defences. In response to stress signals, plants synthesize defence proteins and different chemical compounds. These compounds can carry out direct defensive functions, that is, acting like antioxidants, antibacterial or antifungal. Phenolic compounds and alkaloids, as well as some volatile organic compounds (VOCs) with less polarity and molecular weight, belong to this defensive compounds group. In this Master¿s degree final project we propose the study of the function of a new monoterpene synthase (MTS1) and a salicylic acid 5-hydroxylase (S5H) in tomato plants defensive response against Pseudomonas syringae pv. tomato bacteria. Both genes are involved in plants secondary metabolism. MTS1 takes part in the production of monoterpenic VOCs, such as alpha-terpineol, whose application produces stomatal close. On its behalf, S5H takes part in salicylic acid (SA) degradation to gentisic acid (GA), both defensive phenolic compounds. In the first part of the project, we have observed that MTS1 overexpression produces an increase in some monoterpenes emission which are capable of producing stomatal closing in neighbour non-transgenic plants. Due to one of the possible entry sites for Pseudomonas syringae pv. tomato are stomata, we have evaluated the possible resistance of these transgenic plants, as well as their neighbour plants, against bacterial infection. However, the highest emission of monoterpenes shown is not enough to induce more resistance against the bacteria, neither in transgenic plants nor non-transgenic plants. Our results suggest that terpenoids play a function in interplant communication, doing of overexpression transgenic plants MTS1 a future biotechnological tool. Regarding the S5H role in defensive response, we have observed that a correlation between their expression and their product accumulation exists, the GA, in infected tomato plants with different pathogenic agents. Likewise, silencing transgenic lines in this gene (RNAi_S5H) shows higher SA basal levels than control plants. On the other hand, infected RNAi_S5H transgenic plants with Pseudomonas syringae pv. tomato, exhibit a decrease in GA levels and higher resistance against bacterial infection, confirming the in vivo activity of this enzyme and their function in defensive response. Ultimately, we have seen an advanced senescence phenotype in transgenic plants, suggesting that SA catabolism plays an important role in plants senescence.A lo largo de la evolución, las plantas han ido desarrollando sistemas de defensa frente a diversas agresiones abióticas y bióticas por parte de su entorno. Estos sistemas defensivos incluyen tanto barreras constitutivas como defensas inducibles. En respuesta a las señales de estrés, las plantas sintetizan proteínas de defensa y compuestos químicos de diversa naturaleza. Estos compuestos pueden ejercer funciones defensivas directas, esto es, actuando como antioxidantes, antibacterianos o antifúngicos, o actuar como metabolitos defensivos indirectos, señalizando la respuesta defensiva. Los compuestos fenólicos y los alcaloides, así como algunos compuestos orgánicos volátiles (VOCs) de menor polaridad y peso molecular, pertenecen a este grupo de compuestos defensivos. En el presente Trabajo de Fin de Máster nos planteamos el estudio de la función de una monoterpeno sintasa (MTS1) y de una salicilato 5-hidroxilasa (S5H) en la respuesta defensiva de plantas de tomate frente a la bacteria Pseudomonas syringae pv. tomato. Ambos genes participan en el metabolismo secundario de las plantas. MTS1 interviene en la producción de VOCs monoterpénicos, tales como el alpha-terpineol, cuya aplicación exógena produce el cierre estomático. Por su parte, S5H participa en la degradación del ácido salicílico (SA) a ácido gentísico (GA), siendo ambos, compuestos fenólicos defensivos. En la primera parte del trabajo, hemos observado que la sobreexpresión de MTS1 produce un aumento en la emisión de varios monoterpenos capaces de producir el cierre estomático en plantas vecinas no transgénicas. Puesto que una de las posibles vías de entrada de la bacteria Pseudomonas syringae pv. tomato son los estomas, hemos evaluado la posible resistencia de estas plantas transgénicas, así como de sus vecinas, frente a la infección bacteriana. Sin embargo, la mayor emisión de monoterpenos observada no es suficiente para producir una mayor resistencia a la bacteria, ni en las propias plantas transgénicas, ni en las no transgénicas. Nuestros resultados sugieren que los terpenoides desempeñan una función en la comunicación entre plantas, haciendo de las plantas transgénicas que sobreexpresan MTS1 una futura herramienta biotecnológica. Con respecto al papel de S5H en la respuesta defensiva, hemos visto que existe una correlación entre su expresión y la acumulación de su producto, el GA, en plantas de tomate infectadas con diferentes agentes patogénicos. Asimismo, líneas transgénicas de silenciamiento en dicho gen (RNAi_S5H) presentan unos niveles basales de SA superiores a las de sus correspondientes plantas control. Por otra parte, las plantas transgénicas RNAi_S5H infectadas con la bacteria Pseudomonas syringae pv. tomato, presentan una disminución en los niveles de GA y una mayor resistencia a la infección bacteriana, confirmando así la actividad in vivo de este enzima y su función en la respuesta defensiva. Por último, hemos observado un fenotipo de senescencia avanzada en estas plantas transgénicas, sugiriendo que el catabolismo del SA desempeña un papel importante en la senescencia de las plantas.Minguillón Campos, S. (2018). Papel de MTS1 y S5H en la respuesta defensiva de plantas de tomate frente a la infección con la bacteria Pseudomonas syringae. http://hdl.handle.net/10251/97551TFG

A pregação de Jesus como paradigma para a pregação contemporânea: reflexões exegéticas do Sermão do Monte

The objective of this article is to do an exegetical reading of the Sermon on the Mount, according to the Matthew perspective, looking to see how Christ explained, illustrated and applied his teachings in this pericope, in order to propose parameters for contemporary preachers who want to be inspired by the communication of the Master for the purpose of preaching the Word considering the challenges of the Information society.O objetivo deste artigo é fazer uma leitura exegética do Sermão do Monte, conforme a perspectiva mateana, procurando enxergar como o Cristo explicava, ilustrava e aplicava seus ensinamentos na referida perícope, a fim de propor parâmetros para pregadores contemporâneos que desejam se espelhar na comunicação do Mestre com o propósito de pregar a Palavra frente aos desafios da sociedade da Informação.

As parábolas de Jesus na tradição sinótica: chaves para as ilustrações na pregação contemporânea

The objective of this article is to analyze Jesus' parables in the Synoptic Gospels, particularly their features and purposes, in order to propose parameters for contemporary preachers who wish to be inspired by the Master's communication style for the purpose of improving their sermon illustrations considering the challenges of the information society.O objetivo deste artigo é analisar as parábolas de Jesus conforme registrado nos evangelhos sinóticos, verificando suas características e propósitos, a fim de propor parâmetros para pregadores contemporâneos que desejam se espelhar na comunicação do Mestre com o objetivo de aprimorar suas ilustrações do sermão frente aos desafios da sociedade da Informação