Tesis sobre el tema "Peruvianum"

Lepidium peruvianum Chacón (maca), es una crucífera altoandina, que crece a entre los 3,500 y 4500 m.s.n.m. Originaria de la meseta del Bombón, en los departamentos de Junín y Pasco; por sus cualidades medicinales y su alto valor nutritivo, es una planta de alto interés económico, cuyo cultivo se ha extendido a otras regiones de nuestro país. En el presente trabajo se estudió la susceptibilidad de la especie a las técnicas del cultivo de tejidos como herramienta de producción de metabolitos secundarios. Se realizó la inducción de callos en diferentes explantes de L. peruvianum utilizando la auxina 2,4-D y la citoquinina Kinetina, en un factorial de medios con diferentes concentraciones auxina/citoquinina. Se obtuvieron callos en la mayoría de los medios usados, la relación de hormonas más eficiente fue 1 _M auxina/citoquinina. Se evaluó la presencia de glucosinolatos y alcaloides en los callos obtenidos y se compararon con muestras control de hipocótilos de maca. Se observó la presencia variable de dos fracciones de glucosinolatos en los callos, en la mayoría de los casos las manchas tuvieron una coloración más intensa en los callos que en los controles. De otro lado se observó una alta variabilidad en la presencia de alcaloides y otros metabolitos no identificados en los callos obtenidos en este trabajo. También se evaluó cualitativamente la presencia de mirosinasas en los callos obtenidos, observándose bandas positivas en los callos y las muestras de plantas de maca.
Lepidium peruvianum Chacón, is a Cruciferae native from the Andes. It grows between 3,500 and 4500 m. Original from the Bombón plateau, located at the Peruvian localities Junín and Pasco. It became in a crop with a high economical value, due its medicinal and nutritional properties. Actually, it is extended to other regions of the country. The main objective of this research is to study the tissue culture ability of the crop to use in vitro tissues as a tool for secondary metabolite production. Leaves, petioles, roots and hypocotils of L. peruvianum were tested as explants to induce calli. Different concentrations of 2,4-D and Kinetin, in MS basic medium were tested. Calli were induced in most of the media tested, the most efficient hormone ratio auxin/citokinin was 1. It was evaluated the presence of glucosinolates and alkaloids in the callus induced and compared to maca hypocotils as control sample. Two glucosinolates fractions were obtained from calli analyzed. It was found one or two fractions according to the callus and in most of the cases the concentration was higher in callus than in control. In the other hand, it was observed a high variability in the alkaloid fractions and other unidentified metabolites extracted from the calli evaluated in this work. It was also evaluated the presence of myrosinases in the calli studied, and it was found positive bands either in callus as in maca hypocotils.
Tesis

La entrada PI 126944 de S. peruvianum ha sido descrita como resistente a distintos estreses bióticos, entre ellos a la enfermedad del rizado amarillo del tomate (Tomato yellow leaf curl disease, TYLCD). En trabajos previos realizados en el Instituto Universitario para la Conservación y Mejora de la Agrodiversidad Valenciana (COMAV) se obtuvieron tres híbridos interespecíficos entre esta entrada y el tomate cultivado, empleando el rescate de semillas inmaduras. En trabajos posteriores se desarrollaron generaciones más avanzadas a partir de estos materiales, incluyendo hasta las generaciones pseudo-F6 y retrocruces hacia la especie cultivada de algunas de las generaciones pseudo-Fn. Con objeto de aprovechar estos materiales en la mejora del tomate cultivado se inició, en trabajos anteriores del grupo, el desarrollo de un conjunto de líneas de introgresión (Introgression lines, ILs) que contengan el genoma de la entrada PI 126944 en el fondo genético de S. lycopersicum. Para tener correctamente representado el genoma de la especie donante en el fondo genético del tomate es necesario obtener un gran número de descendientes, por lo que es necesario optimizar los protocolos. El primer objetivo de esta tesis fue el estudio de distintos factores implicados en la superación de las barreras de incompatibilidad, con objeto de conseguir un elevado número de descendientes y maximizar la probabilidad de introgresar el genoma de PI 126944 en el conjunto de ILs. Se llevaron a cabo tres ensayos de retrocruce, iniciados con diferentes plantas de las generaciones pseudo-F4, pseudo-F5 y pseudo-F6. El genotipo de los parentales resultó ser un factor determinante en la obtención de descendencia, por lo que se recomienda utilizar el máximo número de genotipos distintos, con objeto de maximizar la probabilidad de obtener progenie. Se llevó a cabo una prueba de medios de cultivo que permitió seleccionar la combinación más adecuada de auxinas y citoquininas para el cultivo de semillas inmaduras y posterior regeneración de plantas. También se estudió el efecto del número de días transcurridos desde la polinización hasta la recolección de los frutos. Tal y como se ha comentado anteriormente, esta colección del ILs presenta interés por la resistencia del parental donante a TYLCD. Una vez se complete la colección, se evaluará por su resistencia a esta enfermedad. Los estudios previos han mostrado que se trata probablemente de una resistencia poligénica, cuya expresión a menudo es más dependiente de las condiciones ambientales y de otros factores. En estos casos, el empleo de un solo testigo resistente y otro susceptible no es suficiente para determinar con exactitud el nivel de resistencia de los materiales en estudio. La disponibilidad de controles con distintos niveles de resistencia permitiría realizar un fenotipado más preciso. Un grupo de investigadores del "Volcani Center" desarrollaron una escala compuesta por siete líneas con diversos niveles de resistencia a TYLCD, introgresada a partir de distintas especies silvestres. En el marco de esta tesis se evaluó el comportamiento de esta escala en siete ensayos distintos, cuatro de ellos llevados a cabo en el COMAV-UPV y los otros tres en el Instituto Nacional de Ciencias Hortícolas de Cuba. Se compararon dos aislados de TYLCV (Tomato yellow leaf curl virus), una de las especies causantes de TYLCD, diferentes métodos de inoculación, dos estados de desarrollo de la planta y distintas condiciones ambientales. Los síntomas fueron más leves que los obtenidos en las condiciones ensayadas por los autores que desarrollaron la escala, si bien el orden de las líneas en la escala se mantuvo, con pocas excepciones. Sin embargo, en la mayor parte de las condiciones, la respuesta obtenida no correspondió a una escala gradual, ya que se observó una gran diferencia entre los valores de síntomas de las líneas susceptibles y las resistentes, siendo muy pequeñas las difer
Accession S. peruvianum PI 126944 has been reported as resistant to several biotic stresses, among them to Tomato yellow leaf curl disease, (TYLCD). In previous works carried out in the "Instituto Universitario para la Conservación y Mejora de la Agrodiversidad Valenciana" (COMAV) three interspecific hybrids between this accession and cultivated tomato were obtained, using immature seeds rescue. In subsequent works more advances generations from these materials were developed, including until the pseudo-F6 generations and backcrosses to the cultivated species of some of the pseudo-Fn generations. With the aim of taking advantage of these materials in the improvement of cultivated tomato, in previous works of the group, the development of a set of Introgression lines (ILs) that contain the genome of the accession PI 126944 in the genetic background of S. lycopersicum was initiated. The obtaining of a large number of descendants is necessary in order to have correctly represented the genome of the donor species, thus there is the need to optimize the protocols In this sense, the first aim of this work was the study of several factors involved in the overcoming of the incompatibility barriers, to obtain a high number of descendants and maximize the probability of introgressing the genome of PI 126944 in the set of ILs. Three backcross assays were carried out, initiated with different plants of the generations pseudo-F4, pseudo-F5 and pseudo-F6. The parental genotype resulted a determinant factor in the obtaining of descendants, so the recommendation is the use of the maximum number of different genotypes, in order to maximize the probability of obtaining progeny. A proof of different culture media was carried out, which allowed the selection of the better combination of auxins and cytokinins to grow immature seed and regenerate plants. The effect of the number of days from pollination to fruit harvest was also studied. As mentioned before, the collection of ILs results of interest, given the resistance of the donor parental to TYLCD, among others traits. Once the collection is completed, it will be evaluated for its resistance to this disease. The previous studies showed that it is probably a quantitative resistance, whose expression is often more dependent of the environmental conditions and of other factors. In these cases, the use of only one resistant and one susceptible control is not sufficient to determine with accuracy the level of resistance. Availability of controls with different levels of resistance would allow a more precise phenotyping. A group of researchers of the "Volcani Center" developed a scale that consisted of seven lines with different levels of resistance to TYLCD, introgressed from different wild species. In this work, the response of this scale in seven distinct assays was evaluated, four of them carried out in the COMAV-UPV and the others three in the "Instituto Nacional de Ciencias Hortícolas de Cuba". Two isolates of TYLCV (Tomato yellow leaf curl virus), one of the species causing TYLCD, different inoculation method, two development stages of the plant and several environmental conditions were evaluated. In general, the symptoms were more slight that the obtained in the conditions assayed by the authors that developed the scale, although the order of the lines in the scale were maintained, with few exceptions. However, in most of the conditions, the response obtained did not correspond to a graded scale, since there were high differences among the symptom scores of the susceptible and the resistant lines, with very small the differences among the resistant lines. These results suggest the possibility to reduce the number of lines to employ in each concrete condition, selecting the most susceptible, the most resistant and another with intermediate levels of resistance.
L'entrada PI 126944 de S. peruvianum ha sigut descrita com a resistent a diferents estresses biòtics, entre ells a la malaltia de l'arrissat groc de la tomaca (Tomato yellow leaf curl disease, TYLCD). En treballs previs realitzats en el "Instituto Universitario para la Conservación y Mejora de la Agrodiversidad Valenciana" (COMAV) es van obtenir tres híbrids interespecífics entre aquesta entrada i la tomaca cultivada, emprant el rescat de llavors immadures. En treballs posteriors, es van desenvolupar generacions més avançades a partir d'aquests materials, incloent fins a les generacions pseudo-F6 i retrocreuaments cap a l'espècie cultivada d'algunes de les generacions pseudo-Fn. A fi de aprofitar aquests materials en la millora de la tomaca cultivada es va iniciar, en treballs anteriors del grup, el desenvolupament d'un conjunt de línies d'introgressió (Introgression lines, ILs) que continguen el genoma de l'entrada PI 126944 en el fons genètic de S. lycopersicum. Per a tenir correctament representat el genoma de l'espècie donant en el fons genètic de la tomaca és necessari obtenir un gran nombre de descendents, per la qual cosa és necessari optimitzar els protocols. El primer objectiu d'aquesta tesi va ser l'estudi de diferents factors implicats en la superació de les barreres d'incompatibilitat, a fi d'aconseguir un elevat nombre de descendents i maximitzar la probabilitat d'introgressar el genoma de PI 126944 en el conjunt de ILs. Es van dur a terme tres assajos de retrocreuament, iniciats amb diferents plantes de les generacions pseudo-F4, pseudo-F5 i pseudo-F6. El genotip dels parentals va resultar un factor determinant en l'obtenció de descendència, per la qual cosa es recomana utilitzar el màxim nombre de genotips diferents, a fi de maximitzar la probabilitat d'obtenir descendència. Es va dur a terme una prova de medis de cultiu que va permetre seleccionar la combinació més adequada d'auxines i citoquinines per al cultiu de llavors immadures i posterior regeneració de plantes. Es va estudiar l'efecte del nombre de dies transcorreguts des de la pol·linització fins a la recol·lecció dels fruits, havent-se obtingut resultats diferents. Tal com s'ha comentat anteriorment, aquesta col·lecció de ILs presenta interès per la resistència del parental donant a TYLCD. Una vegada es complete la col·lecció, s'avaluarà per la seua resistència a aquesta malaltia. Els estudis previs han mostrat que es tracta probablement d'una resistència quantitativa, l'expressió de la qual sovint és més depenent de les condicions ambientals i d'altres factors. En aquests casos, l'utilització d'un sol control resistent i un altre susceptible no és suficient per a determinar amb exactitud el nivell de resistència dels materials en estudi. La disponibilitat de controls amb diferents nivells de resistència permetria realitzar un fenotipat més precís. Un grup d'investigadors del "Volcani Center" van desenvolupar una escala composta per set línies amb diversos nivells de resistència a TYLCD, introgressada a partir de diferents espècies silvestres. En el marc d'aquesta tesi es va avaluar el comportament d'aquesta escala en set assajos diferents, quatre d'ells duts a terme en el COMAV-UPV i els altres tres en el "Instituto Nacional de Ciencias Hortícolas de Cuba". Es van comparar dos aïllats deTYLCV (Tomato yellow leaf curl virus), una de les espècies causants de TYLCD, diferents mètodes d'inoculació, dos estats de desenvolupament de la planta i diferents condicions ambientals. Els símptomes van ser més lleus que els obtinguts en les condicions assajades pels autors que van desenvolupar l'escala, si ben l'ordre de les línies en l'escala es va mantenir, amb poques excepcions. No obstant açò, en la major part de les condicions, la resposta obtinguda no va correspondre a una escala gradual, ja que es va observar una gran diferència entre els valors de símptomes de les línies susceptibles i les resiste
Campos, GN. (2018). Aprovechamiento de las especies Solanum peruvianum en la mejora del tomate [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/100858
TESIS

El presente trabajo de investigación desarrolla una metodología para la optimización del proceso que incremente la extracción de glucosinolatos principalmente el benciglucosinolato del extracto etanólico de la raíz tuberosa de Lepidium peruvianum Chacón (Maca) bajo la forma de extracto atomizado. El proceso optimizado se inicia con la recolección de la raíz tuberosa de Lepidium peruvianum Chacón (maca) procedente de distrito de San Francisco de Uco (Junín), se le adiciona nitrógeno líquido para disminuir la temperatura a – 50° C que inhibe la actividad de las mirosinasas, enzimas que degradan los glucosinolatos, inmediatamente se procede al molido utilizando un molino de martillos provisto de malla N° 4, luego por malla N° 10, se trasvasa el molido a un tanque de acero con agitador, que contiene alcohol etílico de 80° con agitación constante durante 72 horas a temperatura ambiente; se separa la fase líquida del extracto etanólico utilizando una centrífuga, el líquido separado se transfiere a un tanque enchaquetado, con agitación constante se le adiciona maltodextrina, que es el soporte, se le lleva a una temperatura de 90° C y con el empleo de una bomba se le ingresa al atomizador para la obtención del extracto atomizado en polvo. Se procede a homogenizar utilizando una tamizadora provista de malla N° 60 obteniéndose un polvo fino. La identificación y cuantificación por HPLC del bencilglucosinolato presente en el extracto atomizado utiliza como fase móvil acetonitrilo : metanol : agua (25:25:50) y columna octilsilano 5 um (125 mm x 4.6 mm), el tiempo de retención tanto para el estándar del bencilglucosinolato como para la muestra fue de 9.55 minutos, la cantidad de bencilglucosinolatos en el extracto atomizado del proceso optimizado fue de 0.475 g% , en tanto que para el proceso común fue 0.219 g%. La metodología ofrece una alta especificidad para la extracción de bencilglucosinolato presente en Lepidium peruvianum Chacón (Maca).
The present work of investigation develops a methodology for the optimization of the process that increases the extraction of glucosinolates mainly the benciglucosinolate of the etanolic extract by the tuberous root of Lepidium peruvianum Chacón (maca) under the form of atomized extract. The optimized process begins with the harvesting by the tuberous root of Lepidium peruvianum Chacón (maca) coming from district of San Francisco de Uco (Junín), adds liquid Nitrogen to him to diminish the temperature to - 50° C that inhibits the activity of myrosinases, enzymes which they degrade the glucosinolates, immediately is come to the worn out one using a mill of hammers provided with mesh N° 4, soon by mesh N° 10, it adds the ground one to a steel tank with agitator, who contains etilic alcohol of 80° with constant agitation during 72 hours to room temperature; the liquid phase of the etanolic extract separates using a centrifuge, the separated liquid is transferred to a jacketed tank, with constant agitation adds maltodextrin to him, that is the support, takes to him to a temperature of 90° C and with the use of a pump it is entered to him the spray for the obtaining of the extract atomized in dust. It is come to homogenize using a tamizadora provided with mesh N° 60 obtaining a fine dust. The identification and quantification by HPLC of the bencilglucosinolates present in the atomized extract use like movable phase acetic nitrile: metanol: water (25:25:50) and octilsilane column 5 um (125 mm x 4,6 mm), the time of retention as much for the standard of the bencilglucosinolate as for the sample was of 9,55 minutes, the amount of bencilglucosinolates in the atomized extract of the optimized process was of 0,475 g%, whereas for the common process he were 0,219 g%. The methodology offers a high specificity for the extraction of bencilglucosinolate present in Lepidium peruvianum Chacón (maca).
Tesis

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O gene Sw-5, membro de uma família multigênica com membros dispersos nos cromossomos 9 e 12 do tomateiro, é o principal gene de resistência a tospovírus utilizado nos programas de melhoramento de tomateiro para a indústria e para mesa. O acesso LA 371 de Lycopersicon peruvianum possui resistência de amplo espectro a tospovírus e estudos de introgressão e análise genética da resistência demonstraram que a resistência desse acesso é determinada por um gene dominante localizado no loco Sw-5 ou em um loco proximamente ligado. Estudos prévios haviam levantado a hipótese de que oligonucleotídeos que anelam nas regiões correspondentes ao início da ORF codificada por Sw-5 e na região 5’ deste gene poderiam amplificar seqüências do loco Sw-5 e de um loco proximamente ligado. Esta hipótese foi confirmada neste trabalho. Utilizando esses oligonucleotídeos foram amplificados e clonados dois genes homólogos ao Sw-5, denominados de homólogo 1 e homólogo 2, a partir de DNA extraído de uma planta resistente a tospovírus denominada EP-1 derivada do acesso LA 371. As proteínas putativas codificadas por esses homólogos apresentam 92,94 % de similaridade diferindo em 82 aminoácidos. A proteína 2 é mais similar à proteína codificada pelo gene Sw-5 (99,28%) sugerindo que o homólogo 2 é responsável pela resistência a tospovírus encontrada em LA 371. Essas seqüências foram transferidas para o vetor binário pBI121 e experimentos de transformação encontram-se em andamento visando à comprovação da função desses homólogos na resistência de LA 371 a tospovírus.
The gene Sw-5, member of a multigenic family with members dispersed in tomato chromosomes 9 and 12, is the most used tospovirus resistance gene in tomato breeding programs. The Lycopersicon peruvianum accession LA 371 has a broad-spectrum toposvirus resistance. Genetic analysis has demonstrated that a dominant gene located in the Sw-5 locus or in a closely linked locus is responsible for tospovirus resistance of this accession. Prior studies suggested that oligonucleotides with complementary sequences corresponding to the terminal regions of the Sw-5 ORF would be able to amplify sequences from the Sw-5 locus and from one closely linked locus. This hypothesis was confirmed in this work. DNA extracted from a tospovirus resistant plant called EP-1, derived from the accession LA 371, was used as template for PCR amplification. The PCR product was cloned and restriction analysis revealed that two Sw-5 homologous sequences, called homologous 1 and homologous 2, had been cloned. The putative protein codified by these homologous display 92.94% of similarity for each other, differing in 82 amino acids. The protein coded by homologous 2 is more similar to Sw-5 protein (99.28%) suggesting that this homologous is responsible for the LA 371 tospovirus resistance. These sequences were transferred to the binary vector pBI121 and transformation experiments are in progress aiming to test this hypothesis.
Dissertação importada do Alexandria

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As infecções do trato genital são a causa mais frequente de consulta ginecológica (50-70 % das queixas). Acredita-se que todas as mulheres sexualmente ativas já tiveram pelo menos um episódio de vaginose bacteriana e/ou vulvovaginite. O aumento da resistência das bactérias aos antibióticos convencionais tem estimulado intensos esforços para desenvolver novos agentes antimicrobianos eficazes contra essas bactérias e fungos. O objetivo deste estudo é realizar a produção de formas farmacêuticas plásticas de ação tópica dermatológica e vaginal a partir do extrato seco do hipocotiledon, de Lepidium peruvianum CHACON sp. Nov. = Lepidium meyenii WALP., popularmente conhecida como MACA; esta é uma planta herbácea, bienal ou anual; distribuída basicamente nos Andes Centrais do Peru, em altitudes que vão de 3.500 até 4.500 metros acima do nível do mar. Inicialmente foi feita a identificação botânica e a obtenção de extrato aquoso. Foi realizada a pesquisa de toxicidade da planta; através do teste de toxicidade aguda em camundongos com o extrato bruto, realizado por via intraperitoneal e via oral, a toxicidade foi testada até uma dose de 4.500 mg/kg; e posteriormente foi verificada a ação frente a fungos e bactérias patógenas, comprovando se a ação sobre fungos. Ensaios farmacotécnicos revelaram que o extrato se incorpora perfeitamente nas formulações escolhidas. A determinação das características físicoquímicas dos produtos acabados mostrou que a manipulação não altera qualitativa nem quantitativamente o extrato. Estudos preliminares indicam que as perspectivas de sua utilização nas Indústrias, Cosmética e de Medicamentos são ótimas, pois a utilização destas novas formulações leva a uma diminuição do uso de métodos convencionais, que não raro acarretam ações indesejáveis ao organismo humano.

The domesticated tomato Solanum lycopersicum (L.), formerly known as Lycopersicon esculentum is a genetically well-studied crop species with high-density linkage and molecular maps based on crosses done between cultivated tomato and its distant related wild species. Wild tomato species harbor a wealth of resistance to many pathogens that have been introgressed into domesticated tomato for genetic control of diseases and pests and for improvement of many agronomic traits. The wild tomato S. peruvianum (L.) is the source of the Sw-5 gene, characterized and mapped to chromosome 9 of the tomato genome and introgressed into elite tomato germplasm, providing resistance to the tospovirus Tomato spotted wilt virus (TSWV). TSWV has been reported to be a major problem for tomato growers in many parts of the world, which in some cases, has resulted in tomato fields having been abandoned for some time. Additionally, there are reports that new races of TSWV have evolved that overcome Sw-5. TSWV replicates in both, plant cells and in the alimentary canal cells of thrips and then transmitted to plants by this insect acting as a vector. Both, TSWV and thrips have co-evolved to infest and infect more than 1090 plants species in over 100 families, thrips becoming resistant to pesticides and easily escaping by hiding deep in plant parts. World trade has disseminated thrips all over the world and environmental pressures have forced TSWV to recombine its RNA to overcome new resistance.

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The present reseaech paper was made in the District,Province and Region Lambayeque located in northern Perú during the agricultura seasons: 2004-2005, 2005- 2007 and 2006-2007. The objetive was to evaluate the entomo fauna behavior and the This research work is run in the District, Province and Region Lambayeque, Peru during the crop season 2004-2005, 2005-2006 and 2006 - 2007 with the objective of evaluating the behavior of the entomofauna and yield potential native cotton (Gossypium barbadense L Ssp. Peruvianum).) cotton called "country". The crop was installed in June 2004 beginning with a nursery, in an area corresponding to Ferreñafe Sican Museum and its final installation in the field of Sipan Museum in Lambayeque, at an altitude of 29 meters, average temperature 22.5 º C, humidity on 75% and an average rainfall of 24 mm. 1.333 plants were installed. Also served to assess the ecotypes of color, material valued for its natural color, with a genetic resource of great value to work rescuing this poor germplasm, which is in danger of extisiòn due to the introduction of commercial hybrid , that would put in risk so precious ancestral legacy. We found significant genetic variation in all attributes evaluated. Ferreñafe province had the highest genetic variability in fiber color, the colored ecotypes present five out of nine. The most critical phases of pest attack were sprouting, fruiting and capsule opening. There was a low infestation of worm borer leaf Bucculatrix thurberiella, "Arrebiatadas" Dysdercus peruvianus "Peruvian beaked" Anthonomus vestitus and "wart mite" Acaliptus gossypii, mainly due to unsuitable conditions for their development. The native cotton crop will be greatly affected by the attack of pests if not done an adequate integrated pest management. The more promising ecotypes were: Orange-face, Fine red, white and pink White simple, 2903, 1880, 1642 and 1.24 kilos per plant, respectively. For weight of weed, highlights the ecotypes: Fine Colorado Claro Fifo 3.95 to 2.96 g, respectively, was found in the first harvest of the arboretum A , a greater number of fruit branches with 43,797 branches, with respect to the soca harvest, which formed only 10,643 fruit branches. In the arboretum "B", the results were the same trend. Sample sizes were adequate and representative of the population. The data had a normal or approximately normal distribution.
El presente trabajo de investigación se ejecutó en el Distrito, Provincia y Región Lambayeque-Perú, durante las campañas agrícola 2004-2005, 2005-2006 y 2006- 2007 con el objetivo de evaluar, el comportamiento de la entomofauna y el potencial de rendimiento de algodones nativos (Gossypium barbadense L. Ssp. Peruvianum) , denominado algodón del país . El cultivo se instaló en Junio 2004 iniciándose con un vivero, en un área correspondiente al Museo Sicán de Ferreñafe y su instalación definitiva en el ámbito del Museo de Sipán, en Lambayeque. Se instalaron 1,333 plantones. Además sirvió para evaluar los ecotipos de color, material valioso por su coloración natural, siendo un recurso genético de gran valor para trabajos de rescate de este escaso germoplasma, que se encuentra en peligro de extinción debido a la introducción de híbridos comerciales y cultivares transgénicos, que pondrían en riesgo tan preciado legado ancestral. Se encontró variabilidad genética significativa en todos los atributos evaluados. La provincia de Ferreñafe presentó la mayor variabilidad genética en color de fibra, al presentar cinco ecotipos coloreados de un total de nueve. Las fases más críticas para el ataque de plagas fueron en brotamiento, fructificación y apertura de cápsulas. El cultivo de algodón nativo se verá muy afectado por el ataque de plagas si no se realiza un adecuado manejo integrado de plagas. Los ecotipos más rendidores fueron: Anaranjado-Ante, fino colorado, blanco rosáceo y blanco simple, con 2.903, 1.880, 1.642 y 1.24 kilogramos por planta, respectivamente. Para peso de mota, sobresalieron los ecotipos: Fino colorado y fifo claro con 3.95 a 2.96 g, respectivamente, se encontró en la primera cosecha del arboretum A , en mayor número de ramas fruteras con 43.797 ramas, con respecto a la cosecha soca , que solo formó 10.643 ramas fruteras. En el arboretum B , los resultados tuvieron la misma tendencia.

A regeneração in vitro é bastante utilizada em processos biotecnológicos. No entanto, pouco se conhece sobre os mecanismos envolvidos na aquisição de competência para formação de novos órgãos. Em tomateiro (S. lycopersicum), a alta capacidade de regeneração in vitro é atribuída ao alelo Rg1, vinda de Solanum peruvianum, e que está presente na cv MsK (S. lycopersicum x S. peruvianum). Os genes de nanismo da cv Micro-Tom (MT) foram passados para MsK, obtendo-se, após 8 gerações de autofecundação (F8), a cv Micro-MsK. No presente estudo, após 6 gerações de retrocruzamentos (BC6Fn), criou-se MT-Rg1, com Rg1 isogênico a MT. Testes de regeneração in vitro mostraram que a formação de gemas caulinares adventícias para MT-Rg1 e Micro-MsK são equivalentes, evidenciando que a alta capacidade de regeneração de Micro-MsK é basicamente devido ao Rg1. Plantas MTRg1 apresentam freqüente formação de 3 cotilédones, excesso de ramificações caulinares e senescência atrasada, características ausentes nos parentais. Esses efeitos pleiotrópicos de Rg1 coincidem com aqueles descritos para alterações no hormônio citocinina. Testes de sensibilidade à citocinina e de tempo de senescência realizados por nós, bem como dosagem desse hormônio feita por outras equipes, descartaram essa hipótese. Enxertias recíprocas evidenciaram que a promoção da ramificação caulinar por Rg1 não é um sinal translocável, mas parece ser uma característica intrínseca do tecido. Rg1 também promoveu uma maior formação de raízes adventícias em estacas ex vitro, sugerindo que ele afeta o processo de competência para formação de órgãos em geral, e não somente a indução específica de gemas caulinares. Duplos mutantes entre Rg1 e mutantes com alteração na sensibilidade/metabolismo hormonal (dgt, brt e pro), ausência de ramificações caulinares (ls) e senescência acelerada (l) foram obtidos e testados quanto ao padrão de ramificações e à capacidade de regeneração in vitro. Além desses parâmetros, a presença de Rg1 suprimiu o fenótipo de folhas pouco recortadas do mutante hipersensível a giberelina (pro) e recuperou o sistema radicular pouco desenvolvido do mutante com baixa sensibilidade à auxina (dgt). Rg1 também foi capaz de reverter a ausência de ramificações laterais de ls, sendo que a mutação Me, o qual representa uma superexpressão de um gene do tipo KNOX, não foi capaz. Esse resultado sugere que Rg1 não é um gene do tipo KNOX, embora esses genes sejam considerados os principais controladores da competência. Analisando diferentes tipos de explantes em experimentos independentes, contatou-se que, em geral, as mutações brt (baixa sensibilidade a citocinina), dgt e ls diminuíram a capacidade de regeneração de Rg1, enquanto l aumentou. Rg1 mostrou-se particularmente epistático à mutação pro, revertendo o fenótipo de baixa formação de gemas caulinares desse mutante. Surpreendentemente, Rg1 provou ser mais sensível à auxina em testes de alongamento de segmentos de hipocótilos, sendo capaz de reverter o fenótipo do mutante dgt nesse mesmo teste. Em conjunto, esses resultados confirmam o papel de Rg1 na fase de aquisição da competência e sugerem uma interação dos hormônios giberelina e auxina nesse processo.
The In vitro regeneration process is widely used in plant biotechnology. However, the mechanisms involved in the acquisition of competence for organ formation are hitherto unknown. In tomato (S. lycopersicum), the high capacity for in vitro regeneration is attributed to the Rg1 allele from S. peruvianum, which is present in the cv MsK (S. lycopersicum x S. peruvianum). The dwarfism genes of the cv Micro-Tom (MT) were transferred to MsK, and, after 8 generations of selfing (F8), the cv Micro-Msk was obtained. Here, after 6 generations of backcrosses (BC6Fn), we created the MT-Rg1, which has Rg1 isogenic to MT. Tests of in vitro regeneration showed that shoot formation in MT-Rg1 and Micro-MsK are equivalent, suggesting that the high capacity of regeneration of Micro-MsK is basically due to Rg1. Comparing MT-Rg1 with the control MT, we noticed a high frequency of 3 cotyledon formation, increased shoot branching and late senescence, which are absent in the parentals. These pleiotropic effects of Rg1 coincide with those described for plants with alterations in the hormone cytokinin. Tests of sensitivity for cytokinin and senescence behavior carried out by us, as well as dosage of that hormone made by other researches, discarded that hypothesis. Reciprocal grafting showed that the promotion of shoot branching by Rg1 is not a transmissible signal, but seems to be an inherent characteristic of the tissue. Rg1 also increases adventitious roots formation of ex vitro cuttings, suggesting that it affects the process of competence, which is common for shoots and roots, instead of the specific induction of shoots. Double mutants between Rg1 and mutants with alteration in the sensitivity/metabolism of plant hormones (dgt, brt and pro), as well as mutants with absence of shoot branching (ls) and accelerated senescence (l) were obtained and tested for the capacity of in vitro regeneration. The presence of Rg1 suppressed the phenotype of less dissected leaves of the mutant hypersensitive to gibberrellin (pro) and rescued the phenotype of poor developed root system of the mutant with low auxin sensitivity (dgt). Rg1 was also able to revert the absence of axillary shoot formation in ls, whereas the mutant Me, which represents an over expression of a KNOX gene was not. This result suggests that Rg1 is not a KNOX gene, although those genes are considered the main controllers of the competence. Analyzing different types of explants in independent experiments, it was verified that, in general, the mutations brt (low sensibility to cytokinin), dgt and ls decreased the regeneration capacity of Rg1, while l increased it. Rg1 was particularly epistatic to the pro mutation, reverting the phenotype of little shoot formation of that mutant. Surprisingly, Rg1 proved to be more sensitive to auxin in the hypocotyl segment elongation test, being able to revert the phenotype of the mutant dgt in the same test. Together, these results confirm the role of the Rg1 in the process of acquisition of the competence and suggest an interaction of the hormones gibberellin and auxin in this process.

Among viral diseases affecting cultivated tomato, Tomato yellow leaf curl disease (TYLCD) is one of the most devastating. This disease is caused by a complex of viruses of which Tomato yellow leaf curl virus (TYLCV) is regarded as the most important species. Current control strategies to fight viral diseases in tomato are mainly based on genetic resistance derived from wild relatives. In the present thesis, resistance derived from S. chilense and S. peruvianum has been exploited in breeding for resistance to TYLCD. In a previous study, TYLCV-resistant breeding lines derived from LA1932, LA1960 and LA1971 S. chilense accessions were developed. Therefore, the first objective of this thesis was to study the genetic control of the resistance derived from these accessions. With this aim, response to viral infection was assayed in segregating generations derived from the aforementioned resistant lines. The results obtained were compatible with a monogenic control of resistance. Resistance levels were higher in LA1960- and LA1971-derived F2 generations, as shown by slighter symptoms in the resistant plants and a higher number of asymptomatic plants compared with the results obtained in the LA1932-derived F2 generation. It is noteworthy that the level of resistance present in our materials is comparable to or even higher than the levels found in tomato lines homozygous for Ty-1. The response in plants heterozygous for the resistance gene was comparable to the response in homozygous plants for all three sources employed. This implies that the resistance genes derived from all three sources seem to be almost completely dominant. This effect was stronger for LA1971-derived resistance. The results were similar when comparing viral accumulation, as was expected, since a positive correlation was found in these families between viral accumulation and symptom scores. This has important implications in breeding, since the resistance will be used mostly for hybrid development. Our second objective was to map the loci associated with the major resistance genes identified. A total of 263 markers were screened, 94 of them being polymorphic between both species. Recombinant analysis allowed the resistance loci to be localized on chromosome 6, in a marker interval of 25 cM. This interval includes the Ty-1/Ty-3 region, where two S. chilense-derived TYLCD resistance loci were previously mapped. In order to test if the resistance genes identified in our populations were allelic to Ty-1 and Ty-3, further fine mapping was carried out. A total of 13 additional molecular markers distributed on chromosome 6 allowed 66 recombinants to be identified, and the resistance region to be shortened to a marker interval of approximately 950 kb, which overlaps with the Ty-1/Ty-3 region described previously by other authors. Therefore, the results obtained indicate that closely linked genes or alleles of the same gene govern TYLCV resistance in several S. chilense accessions. The third objective of the present thesis was to start the construction of a set of introgression lines (ILs) derived from Solanum peruvianum accession PI 126944 into the cultivated tomato genetic background. Once this collection of ILs is developed, it will represent a powerful tool for exploiting the resistance to different pathogens found in this particular accession in addition to other possible characters of interest. The starting plant material consisted of several segregating generations that were derived from two interspecific hybrids previously obtained by our group. Many crosses and embryo rescue were required to obtain subsequent generations due to the high sexual incompatibility that exists between tomato and PI 126944. Several mature fruits from the most advanced generations produced a few viable seeds, although embryo rescue was also employed to obtain progeny. As only a few plants were obtained by direct backcrossing, additional crosses were made in order to increase the number of descendants. A high degree of incompatibility was also found in crosses between sib plants. A total of 263 molecular markers were tested in some generations, 105 being polymorphic between tomato and PI 126944. Available generations were genotyped with these polymorphic markers in order to determine which alleles of S. peruvianum were already introgressed. On average, 79, 78 and 84 % of the S. peruvianum genome was represented in the pseudo-F2, pseudo-F4 and pseudo-F5 generations, respectively, for the markers analyzed. A reduction in the S. peruvianum genome was observed in more advanced generations, such as BC1 (56 %), pseudo-F2-BC1 (60 %) and pseudo-F3-BC1 (70 %). A greater reduction was observed in the pseudo-F3-BC2 generation (33 %). As a consequence of the reduction in the S. peruvianum genome, a loss of incompatibility was observed in some cases. The S. peruvianum genome was almost completely represented among the different plants of the most advanced generations. An evaluation for resistance to TYLCD and Tomato spotted wilt virus (TSWV) was carried out in some of the advanced generations, some of which were resistant to one or both viruses. In conclusion, we have conducted a successful and deeper exploitation of two wild species with proved resistance to TYLCD, S. chilense and S. peruvianum, identifying and fine mapping new genes of resistance.
Julián Rodríguez, O. (2014). Exploitation of Solanum chilense and Solanum peruvianum in tomato breeding for resistance to Tomato yellow leaf curl disease [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/36867
TESIS

Oritrophium peruvianum (Lam.) Cuatr. is a perennial species, endemic to the Páramos of the Cordillera of Los Andes. In Venezuela, where it is known as Frailejón morado, it is harvested for its widely acclaimed medicinal properties in treating asthma, bronchitis, and influenza. This study aimed to investigate the biology and ecology of O. peruvianum and the impacts of harvesting on its populations. Undisturbed populations of O. peruvianum were dominated by adult vegetative individuals, with middle-sized plants prevailing and with low densities of large adults, whilst densities of small plants were variable. The population structure changed slightly after one year and annual plant growth was shown to be extremely slow. Flowering started at the beginning of the rainfall period and overlapped with fruiting throughout the wet season (April to November), with only a few scattered individuals reproducing during the dry season. Seed production was very variable and germination under experimental conditions was successful (51 to 88%). The population dynamics of two populations were analyzed using a five stages time-invariant linear matrix model with field data collected over two years. The growth rate of the first population was higher (?1 =1.32) than that of the second population (?1 = 1.13). Sensitivity analysis showed that their growth rates were more sensitive to a change in the survival of individuals than to reproduction, due to the low survival rate of newly emerged seedlings. Plants were contributing more to the growth rate if they remained in the same stage class than if they grew to the following stage class. The most significant transition was the survival of stage class 3 (small adults) whilst stage class 5 (large adults) contributed little to the population growth rate. A uniform harvesting of the adult stages each year up to 21% and 43% respectively in each of the two populations could be applied without threatening their survivals. The harvesting of small adults would affect the growth rates of the populations most, whilst the complete removal of stage class 5 would have the least impact. The most sustainable practical policy, considering larger plants with higher economic value, allows harvesting of all plants from stage class 5 in both populations, each year. In the population with the higher proportion of plants in stage class 3, more than one half (63%) of this stage class could be also harvested, whilst in the second population 83% of the plants from stage class 4 (middle-sized plants) could be harvested instead.

Las raíces de Lepidium peruvianum Chacón (maca) son reconocidas por sus propiedades curativas dentro de la medicina tradicional, estas poseen metabolitos secundarios con actividad biológica como alcaloides, flavonoides y glicosinolatos que podrían estimular la recuperación del sistema inmune en animales con inmunosupresión experimental por Ciclofosfamida (CP) que induce supresión del sistema inmune humoral en ratones. El presente trabajo tuvo como objetivo comprobar el efecto modulador de la maca sobre la respuesta inmune humoral en ratones suprimidos con ciclofosfamida. Fueron obtenidos tres tipos de extractos: clorofórmico (ECl), metanólico (EMe) y acuoso (EAc) que se administraron por vía oral en dosis de 300mg/Kg del peso corporal.
The roots of Lepidium peruvianum Chacón (maca) they are recognized by theirs medicine properties inside the traditional medicine. They have secondary metabolites with biological activity as alkaloids, flavonoids and glucosinolates that can stimulates the recovery of the immune system in animals with experimental immunosuppretion by cyclophosphamide (CP) that induces suprresion of the immune system humoral in mice. The objective of the present study was verify the modulating effect of the “maca” on the immune response humoral in mice suppressed with cyclophosphamide. Three types of extracts were obtained: chloroformic (ECl), methanol (EMe) and aqueous (EAc) that they were administered for oral route in dose of 300mg/Kg of the body weight.
Tesis

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Plantas dos acessos de Lycopersicon peruvianum (LA 444/1-19, LA 371-20, PI 126444-3, PI 126928-2, PI 126944-6 e PI 126944-12) e de L. chilense (LA 130-5 e LA 2753-11) que possuem resistência de amplo espectro a tospovírus foram utilizadas no cruzamento com a cultivar Moneymaker (L. esculentum) para produção de híbridos interespecíficos, visando a introgressão dessa resistência no tomateiro cultivado. Para superar as barreiras interespecíficas verificadas nestes cruzamentos utilizou-se o cultivo in vitro de sementes imaturas em meio de cultura Murashige & Skoog (MS). Obtiveram-se cinco híbridos do cruzamento L. esculentum x LA 130-5, quatro do cruzamento L. esculentum x LA 2753-11, quatro do cruzamento L. esculentum x LA 371-20, cinco do cruzamento L. esculentum x LA 444/1-19, um do cruzamento L. esculentum x PI 126944-6 e um do cruzamento L. esculentum x PI 126944-12. Não se obteve nenhuma planta dos cruzamentos entre L. esculentum e os acessos PI 126444 e PI 126928, mesmo utilizando-se o cultivo in vitro de semente imatura. Todos os híbridos foram auto-incompatíveis e apresentaram características morfológicas intermediárias aos seus progenitores. A natureza híbrida das plantas F 1 foi confirmada utilizando-se um marcador molecular CAPS. Alguns dos híbridos foram propagados mediante o processo de estaquia e inoculados para avaliação da resistência com isolados de tospovírus, pertencentes às espécies Tomato spotted wilt virus - TSWV, Groundnut ringspot virus GRSV e Tomato chlorotic spot virus - TCSV. Os híbridos foram submetidos a duas inoculações via extrato foliar tamponado, realizadas num intervalo de sete dias. Apenas os híbridos do cruzamento L. esculentum x LA 371-20 e L. esculentum x PI 126944-6 foram resistentes aos três isolados utilizados. Os demais híbridos foram resistentes aos isolados das espécies GRSV e TCSV, mas suscetíveis ao isolado da espécie TSWV. Os híbridos foram retrocruzados com a cultivar Moneymaker para obtenção de populações de retrocruzamento (RC). Populações RC 1 para os híbridos L. esculentum x LA 130-5, L. esculentum x LA 2753-11, L. esculentum x LA 371-20 e L. esculentum x LA 444/1-19 foram obtidas utilizando-se também o cultivo in vitro de sementes imaturas, obtendo- se um total de 99 plantas RC 1 . Estas plantas foram autofecundadas e novamente retrocruzadas com a cultivar Moneymaker, gerando populações RC 1 F 2 e RC 2 , respectivamente. O estudo da herança da resistência em uma população RC 1 F 2 derivada de L. peruvianum LA 371-20 indicou que a resistência originária dessa fonte é condicionada por um gene dominante, sendo efetiva contra três espécies de tospovírus (TSWV, TCSV e GRSV). A análise genética dessa progênie com um marcador molecular CAPS ligado ao gene Sw-5 demonstrou que o gene de resistência derivado do acesso LA 371-20 é um alelo do loco Sw-5 ou um gene proximamente ligado.
Accessions of Lycopersicon peruvianum (LA 444/1-19, LA 371-20, PI 126444-3, PI 126928-2, PI 126944-6, PI 126944-12) and L. chilense (LA 130-5, LA 2753-11) that possess wide spectrum tospovirus resistance were used in crosses with the cultivar Moneymaker (L. esculentum) for production of interspecific hybrids, seeking the introgression of this resistance into cultivated varieties. The interspecific barriers in these crossings were overcome by cultivating immature seeds in Murashige & Skoog medium. Hybrids were obtained from crosses ́Moneymaker ́x LA 130-5; ́Moneymaker ́x LA 2753-11; ́Moneymaker ́x LA 371-20; ́Moneymaker ́ x LA 444/1-19; ́Moneymaker ́ x PI 126944-6 and ́Moneymaker ́ x PI 126944-12. Seeds derived from crosses between ́Moneymaker ́and accessions PI 126444 and PI 126928 did not germinated or produced abnormal seedlings. All hybrids were self-incompatible and displayed morphological characteristics intermediate to their progenitors. The hybrid nature of the F 1 plants was also confirmed by molecular marker analysis. Cuttings from some of the hybrids were inoculated with virus isolates, belonging to the species Tomato spotted wilt virus - TSWV, Groundnut ringspot virus - GRSV and Tomato chlorotic spot virus - TCSV. Only the cuttings from ́Moneymaker ́x LA 371-20 and ́Moneymaker ́x PI 126944-6 hybrids were resistant to all isolates. The others hybrids were resistant to GRSV and TCSV, but susceptible to the TSWV isolate. The hybrids were backcrossed to ́Moneymaker ́ for production of backcross populations (RC). RC 1 populations were obtained for ́Moneymaker ́ x LA 130-5, ́Moneymaker ́x LA 2753-11, ́Moneymaker ́ x LA 371-20 and ́Moneymaker ́ x LA 444/1-19. The RC 1 plants were selfed and again backcrossed to ́Moneymaker ́ to obtain RC 1 F 2 and RC 2 populations, respectively. Inheritance studies of the resistance in a population RC 1 F 2 derived from LA 371-20, showed that the resistance of this accession is conditioned by a dominant gene effective against three tospovirus species (TSWV, TCSV and GRSV). Genetic analysis of this progeny with a molecular CAPS marker linked to Sw-5, demonstrated that the gene derived from LA 371-20 is allelic to Sw-5 or a tightly linked gene.
Dissertação importada do Alexandria

La investigación busca medir la producción de anticuerpos anti-HBs al 1, 2 y 6 meses, evaluar la seroconversión y seroprotección en ambos grupos; cultivar células mononucleares de sangre periférica para el estudio de la producción de óxido nítrico (NO), comparar el recuento diferencial de la población de linfocitos CD3+CD4+ (LT cooperadores) y CD3+CD8+ (LT citotóxicos) y finalmente realizar un hemograma a todos los voluntarios al mes y a los 2 meses de iniciado el tratamiento para el recuento de leucocitos, eritrocitos, neutrófilos, linfocitos, basófilos, eosinófilos y monocitos; así como también determinar los niveles de hemoglobina. La Lepidium peruvianum Chacón (Brassicaceae), también conocida como maca, es una planta medicinal que crece exclusivamente entre 4,000 y 4,500 msnm en los Andes del Perú, presenta diferentes ecotipos que varían de acuerdo al color de la raíz, que va desde el blanco al negro; en la actualidad es ampliamente usada por sus importantes propiedades nutritivas y medicinales relacionadas con la presencia de diversos metabolitos secundarios. Para evaluar el efecto del tratamiento con maca sobre la respuesta inmune tanto humoral como celular se inmunizaron 70 personas voluntarias adultas y sanas con la vacuna contra el virus de la hepatitis B, Gene Vac B. Se formaron 2 grupos, uno se trató con maca pulverizada (3g/día) y el control fue tratado con placebo. Se aplicó un estudio aleatorio a doble ciego, todos los participantes recibieron las tres dosis de la vacuna de acuerdo al esquema de vacunación (0, 1, 6 meses) y durante 2 meses contados a partir de la primera dosis ambos grupos recibieron una dosis diaria de maca y placebo respectivamente.
Tesis

Lepidium peruvianum (maca), es un cultivo tradicional de los Andes Centrales del Perú y es empleada desde tiempos precolombinos como una planta medicinal y alimenticia. Los cultivares de maca se diferencian por el color externo de la raíz denominándose a cada uno como ecotipo. Estudios anteriores reportaron que los diferentes ecotipos de maca presentan diferencias en cuanto a su actividad biológica. Los objetivos del presente estudio fueron: Determinar la concentración de flavonoides, calcio y hierro en el extracto metanólico (EM) de los ecotipos blanco, morado, rojo y negro; comprobar la actividad moduladora del EM de los ecotipos blanco, morado, rojo y negro sobre la respuesta inmune humoral in vivo frente a los glóbulos rojos de carnero (GRC) en animales normales e inmunosuprimidos con ciclofosfamida (CF), así como su efecto sobre el peso y celularidad de los órganos linfoides y recuento de las células sanguíneas. Finalmente, determinar la producción de óxido nítrico (ON) por macrófagos peritoneales cultivados con el EM de los ecotipos seleccionados.
--- Lepidium peruvianum (maca) is a traditional crop in the Central Andes from Peru, it is well-known and employee from pre-Columbian times as a medicinal and nutritional plant. Maca is presented in different ecotypes according to colors of its roots. Previous studies reported that the different ecotypes from maca display differences in their biological activity. The objectives of the present study were: To determine flavonoids, calcium and iron concentration in methanolic extract (EM) of white, purple, red and black ecotypes, to verify the modulatory activity of the methanolic extract of the white, purple, red and black ecotypes on the humoral immune response in mice to sheep red blood cells (SRBC) in normal and inmunosuppressed animals with cyclophosphamide (CP), as well as its effect on the weight and cellularity of the lynphoid organs and peripheral blood cell count. Finally to evaluate the activity of the methanolic extracts of the selected ecotypes on the nitric oxide (NO) production by peritoneal macrophages.
Tesis

La infección con el virus de la inmunodeficiencia humana (HIV), causa una profunda inmunodepresión que expone a las personas infectadas a diversas infecciones oportunistas. Lepidium peruvianum Ch. presenta varias propiedades como, su capacidad estimulante de la reproducción y energizante, investigaciones recientes han demostrado su capacidad antitumoral e inmunoestimulante. El objetivo del presente trabajo fue determinar la actividad de tres tipos de extractos (clorofórmico, acuoso y metanólico) de L. peruvianum Ch. en cultivos de leucocitos de personas saludables y HIV-1+. Para ello, células sanguíneas de 5 voluntarios sanos y 5 HIV-1+ fueron cultivadas por triplicado en presencia de cada extracto a una concentración final de 800µg/mL y se incubó por 16 horas a 37 ºC. Se realizó el recuento de los leucocitos y sus 4 estirpes celulares (linfocitos, neutrófilos, eosinófilos y monocitos) mediante hemograma manual y de las poblaciones de linfocitos T CD3+CD4+ y CD3+CD8+ por citometría de flujo. Se empleó el medio de cultivo RPMI como control negativo de la prueba. En las personas saludables, para las 4 estirpes celulares evaluadas, el extracto clorofórmico logró incrementar significativamente el número de linfocitos (p=0.003). La población linfocitaria T CD3+CD4+ presentó un incremento celular con el extracto clorofórmico (p= 0.004).En las personas HIV-1+, los 3 extractos lograron incrementar el número de leucocitos. En los individuos pertenecientes al estadío C se observó que el extracto clorofórmico logró un incremento significativo del número de linfocitos respecto al control. Estos resultados pueden ser de gran ayuda para utilizar a L. peruvianum Ch. en el tratamiento de personas infectadas con HIV-1 contribuyendo con restaurar el nivel y funcionamiento de las células T infectadas por el virus. Palabras Clave: Lepidium peruvianum, Lepidium meyenii, maca, inmunoestimulante, metabolitos secundarios, HIV-1, SIDA.
--- Infection with the human immunodeficiency virus (HIV), causes a profound immunosuppression that exposes people infected to various opportunistic infections. Lepidium peruvianum Ch. has several properties, the most known, its ability stimulating the reproduction and energizing or revitalizing, recent research has shown antitumor and immunostimulatory capacity. The aim of this study was to determine the activity of three types of extracts (chloroform, methanol and aqueous) of L. peruvianum Ch. in cultures of leukocytes from healthy people and HIV-1+. To do this, blood cells from 5 healthy and 5 HIV-1+ volunteers were cultured in triplicate in the presence of each extract at a final concentration of 800μg/mL and incubated for 16 hours at 37 ° C. We performed the counting of leukocytes and their 4 cell lines (lymphocytes, neutrophils, eosinophils and monocytes) through a hemogram and populations of T lymphocytes CD3+ CD4+ and CD3+ CD8+ by flow cytometry. We employed RPMI culture medium as negative control test. In healthy people, for the 4 cell lines tested, the chloroform extract was able to increase significantly the number of lymphocytes (p= 0.003). The CD3+CD4+ lymphocyte population showed an increase cell with the chloroform extract (p= 0.004). In people HIV-1+ the 3 extracts were able to increase the number of leukocytes. In individuals belonging to stage C showed that the chloroform extract achieved a significant increase in number of lymphocytes over control. These results may be helpful to use a L. peruvianum Ch. in treating people infected with HIV-1 contributing to restore the level and function of T cells infected with the virus. Key Words: Lepidium peruvianum, Lepidium meyenii, maca, immunostimulant, secondary metabolites, HIV-1, AIDS.
Tesis

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Alguns aspectos da interação entre o gene de resistência Sw-5 e os tospovírus, foram analisados neste trabalho. A capacidade de Sw-5 conferir resistência em Nicotiana benthamiana foi avaliada em plantas transformadas com uma construção na qual a ORF e a região 3' do gene estavam sob controle do promotor 35S. As plantas transformadas foram resistentes à infecção por tospovírus. A comparação do espectro da resistência destas plantas com o observado para outras solanáceas , indica que as vias de sinalização e as respostas de defesa ativadas por Sw-5 estão conservadas nesta família, e que o polimorfismo genético nos componentes das vias de transdução de sinais pode resultar em diferentes níveis de resistência. A fim de identificar o gene de avirulência dos tospovírus, os genes N, NSm e NSs , foram expressos isoladamente ou em combinações, por meio do vetor viral PVX, em plantas com Sw-5. A expressão destes genes não foi capaz de desencadear a resposta de hipersensibilidade e tampouco interferiu na infecção da planta por PVX. Portanto, outro componente dos tospovírus deve ser responsável pelo desencadeamento da reação de resistência. Independentemente da presença do gene Sw-5, a expressão do gene NSs por meio do PVX , agravou os sintomas provocados por este vírus em algumas solanáceas, o que pode ter relação com a capacidade desta proteína de suprimir silenciamento gênico. Em plantas com Sw-5, a co-expressão da região 5' deste gene por meio do vetor PVX , favoreceu a infecção sistêmica por tospovírus. Uma vez que o efeito foi observado tanto para expressão senso quanto anti-senso, a redução dos níveis de mRNA de Sw-5 provocada por silenciamento gênico , poderia ser a causa desta interferência na resistência, embora a análise de Northern blot não tenha demonstrado tal redução. Uma seqüência homóloga a Sw-5 contendo uma ORF sem deleções ou interrupções prematuras foi clonada a partir do acesso LA371-20 de Lycopersicon peruvianum. Análises moleculares demonstraram que esta seqüência é originada do loco Sw-5 ou de região próxima a este e que segrega com a resistência a tospovírus. A capacidade deste e de outros três homólogos oriundos de distintos acessos de tomateiro de conferir resistência a tospovírus, foi avaliada em plantas transgênicas de tomateiro e tabaco. As plantas transformadas foram suscetíveis ao vírus. A não-funcionalidade destes homólogos pode ser devido à estrutura das construções utilizadas na transformação. Alternativamente, outros homólogos presentes nestes acessos de tomateiro podem ser os responsáveis pela resistência.
In this work some aspects of the tospovirus - Sw-5 interaction was analyzed. The capacity of the Sw-5 to confer resistance in Nicotiana benthamiana was evaluated in transgenic plants transformed with Sw-5 ORF containing its own 3 ́ UTR region under 35S promoter control. Transgenic plants were resistant to tospovirus infection. Comparisons of the resistance spectrum with other members of the Solanaceae suggest that the signal transduction pathways and resistance responses triggered by Sw-5 are conservated in solanaceae and that the genetic polymorphism in the signal transduction components may result in different resistance levels. The N, NSm and NSs genes isolated or in combination were expressed by PVX vector in plants harboring Sw-5 in order to detect the tospovirus avirulence gene. These genes were not able to trigger the hypersensitive response and to affect PVX infection in Sw-5 plants, which suggest that another tospovirus component is the elicitor of the resistance response. Independently of the Sw-5 gene, PVX clones harboring NSs gene induced more severe symptoms in some solanaceae plants. Gene silencing may be the cause of this symptoms. In transgenic plants harboring Sw-5 gene, the co-expression of the 5 ́ region of this gene by PVX favored tospovirus infection. As this effect was observed both for 5 ́ sense and anti- sense constructions it is possible that it has been caused by reduction on mRNA levels by gene silencing, although Northern blot analysis did not agree with this hypothesis. An Sw-5 homolog was cloned from LA371-20 accession. This homolog is localized in or near of Sw-5 locus and segregated with tospovirus resistance. The capacity of this and other three homologs originated from others accessions to confer tospovirus resistance was evaluated in tobacco and tomato transgenic plants. All the transformants were susceptible to the virus. Other homologs presents in the different accessions evaluated may be responsible for the resistance, although problems in the structure of the constructions can not be discarded.
Tese importada do Alexandria

Se determinó la respuesta de resistencia de 10 patrones de tomate a una población avirulenta de Meloidogyne javanica en maceta. Los ensayos se realizaron en primavera, cuando las temperaturas permitían la expresión fenotípica de la resistencia del gen Mi-1 (28˚C), en verano sometidos a altas temperaturas y en campo, exponiéndolos a altas densidades poblacionales del nematodo. A temperaturas inferiores a 28˚C los patrones mostraron gran variabilidad en la respuesta de resistencia que osciló entre alta y moderadamente resistente (PG-76, Gladiator, MKT-410; Brigeor, 42851, 43965, Big Power y He-man), hasta susceptible (Beaufort, Maxifort). Por encima de 28˚C, sólo dos patrones (PG-76 y He-man) inhibieron la reproducción del nematodo. Frente a distintas poblaciones de M. arenaria, M. incognita y M. javanica, el patrón PG-76, fue altamente resistente a todas las poblaciones, Brigeor osciló entre altamente resistente y moderadamente resistente, mientras que Beaufort y Maxifort mostraron menor resistencia o fueron totalmente susceptibles; además ésta varió en función de la población analizada. Se caracterizó molecularmente el locus Mi-1 en los patrones híbridos y cultivares de tomate estudiados. Se emplearon los marcadores moleculares PM3, PMi y Mi23, específicos para la caracterización del locus Mi en patrones híbridos de tomate (S. lycopersicum × S. habrochaites; S. lycopersicum × S. chilense), mediante PCR. También se realizaron análisis bioinformáticos con marcadores específicos (Mint-up/do, C172, C2S4, IMO-F1/R1, y VIGS) para determinar la presencia del gen Mi-1.2 en dichos patrones. Los resultados mostraron que los marcadores PMi y Mi23 amplifican homólogos del gen Mi-1 en S. chilense, S. habrochaites y S. peruvianum y también en S. lycopersicum (marcador Mi23). El marcador PM3 amplificó el gen Mi-1.2 en Beaufort y Maxifort (S. lycopersicum × S. habrochaites) pero no fue efectivo para los híbridos de S. chilense. El marcador molecular PM3, no pudo determinar la expresión de del gen Mi-1.2 en Beaufort y Maxifort por hallarse fuera de la secuencia codificadora (CDS) del gen. Análisis bioinformáticos indicaron que ningún marcador específico diseñado para el gen Mi-1.2, podía este gen de otros homólogos presentes en S. lypcopersicum y S. peruvianum. El nuevo marcador Pau-Do en combinación con el primer C2S4, amplificaron un fragmento de 1.494 pb en la CDS del gen Mi-1.2 en raíces y hojas de Beaufort y Maxifort. La durabilidad de la resistencia del gen Mi-1 después del cultivo reiterado de patrones de tomate se determinó en ensayos de campo durante tres años consecutivos, empleando PG-76 y Brigeor. El patrón PG-76 fue muy resistente después del 1er ciclo de cultivo, pero mostró resistencia intermedia y suscetibilidad al finalizar el 2o y el 3er año de cultivo, respectivamente. El patrón Brigeor y el cultivar de tomate resistente Monika (control) mantuvieron un nivel de resistencia intermedio al final del 3er cultivo, aunque ensayos posteriores confirmaron la aparición de virulencia. Los resultados mostraron que el cultivo reiterado de patrones de tomate resistentes seleccionó rápidamente aislados virulentos de M. javanica. El fenotipo virulento de estas poblaciones se analizó molecularmente con el marcador MVC, diseñado para distinguir las poblaciones virulentas seleccionadas de Meloidogyne de los aislados naturalmente virulentos. Se analizaron dos poblaciones japonesas seleccionadas de M. incognita y M. javanica, tres poblaciones españolas virulentas seleccionadas, una población naturalmente virulenta y una avirulenta (todas M. javanica). Las muestras de ADN se obtuvieron de individuos juveniles o de hembras adultas y se incluyeron muestras de agua sin nematodos
(5 µm filtrada) procedentes del drenaje de una maceta con una planta infectada por una población virulenta japonesa. El marcador MVC amplificó ADN en las muestras de agua pero no en las que sólo contenían ADN de nematodos. Las secuencias de ADN mostraron una estrecha correlación con diversas proteínas de especies de betaproteobacterias. Los experimentos revelaron que el marcador de MVC no está relacionado con un gen de virulencia del nematodo (avr) sino con betaproteobacterias. Finalmente, se estudió la existencia de homólogos del gen Mi en las especies de tomate silvestre Solanum chilense, S. habrochaites, S. peruvianum y S. huaylasense. La respuesta de resistencia de la variedad LA-1358 de S. huaylasense varió en función de la especie del nematodo estudiada: fue resistente frente a M. arenaria y susceptible frente a M. javanica. La reproducción de M. incognita fue muy variable y no difirió de la reproducción alcanzada en los dos cultivares empleados como controles.
The response of 10 Mi-1 tomato rootstocks to a Mi-avirulent population of M. javanica was determined in pot tests conducted in a greenhouse in spring when temperatures remained below the Mi-1 functionality resistance threshold (28 ˚C), and in summer when daily temperatures exceeded the Mi-1 expression threshold. Rootstocks were also evaluated in the field exposing them to high population densities of the nematode. Results on infectivity and reproduction below 28 ˚C indicated a wide variability in the resistance response of the rootstocks ranging from highly or intermediate resistance (PG-76, Gladiator, MKT-410; Brigeor, 42851, 43965, Big Power and He-man) to fully susceptible (Beaufort and Maxifort). At high temperature conditions, only PG-76 and He-man inhibited the reproduction of M. javanica. Rootstocks PG-76, Brigeor, Beaufort and Maxifort were challenged to different populations of M. arenaria, M. incognita and M. javanica. Rootstock PG-76 was highly resistant to all the populations tested, whereas the response of Brigeor ranged from highly to moderate resistance; the resistance response of rootstocks Beaufort and Maxifort varied according to the population tested. Molecular characterization of the resistance phenotype was performed for all the tomato hybrid rootstocks and cultivars tested. The markers PM3, PMi, Mi23, for the characterization of the Mi-locus of hybrid tomato rootstocks (S. lycopersicum × S. habrochaites and S. lycopersicum × S. chilense) were used for PCR reactions. In silico analyses were done with specific markers for the Mi-1.2 gene (Mint-up/do, C1/2, C2S4, IMO-F1/R1, and VIGS). Markers PMi and Mi23 were polymorphic for the Mi-1 locus in wild Solanum species (S. chilense, S. habrochaites, and S. peruvianum) and for S. lycopersicum (marker Mi23). Marker PM3 detected the Mi-1.2 gene in S. lycopersicum × S. habrochaites hybrid rootstocks, but not in the S. chilense hybrids. As marker PM3 is located outside the coding sequence of the Mi-1.2 gene, expression of this homolog could not be determined in Beaufort and Maxifort. In silico results indicated that none of the available markers for the Mi-1.2 gene could distinguish this homolog from the other Mi-homologs from S. lypcopersicum and S. peruvianum species. A new marker Pau-Do, in combination with C2S4, was designed to amplify in CDS of the Mi-1.2 gene. Amplification with these primers of cDNA from Beaufort and Maxifort indicated that the Mi-1.2 gene was expressed in both rootstocks, despite their susceptible phenotypic response to some Meloidogyne populations. The durability of the Mi-1 gene after repeated cultivation of resistant tomato rootstocks (PG-76 and Brigeor) was determined through field trials during three consecutive years. Rootstock PG-76 responded as highly resistant after the first cropping cycle, although it became fully susceptible after the second and the third cropping cycles. Rootstock Brigeor and the resistant tomato cultivar Monika (control), retained intermediate resistance levels at the end of the third year. Bioassays confirmed that selection of virulence occurred more rapidly in plots with rootstock PG-76 followed by Brigeor and the resistant tomato cultivar Monika. The virulent phenotype of the selected M. javanica populations in the field experiments was determined with MVC molecular marker, designed to distinguish selected from naturally virulent populations of Meloidogyne spp. The populations analyzed included two Japanese selected virulent populations, and the three virulent populations selected in the field trials, and one naturally virulent population and one avirulent population from Spanish. DNA samples were obtained from individual juveniles (J2) or adult females from all the selected virulent populations. Experiments included water samples free of nematodes (5-µm filtered), obtained from the draining-water of a plant infected by a Japanese selected virulent population. Amplification of DNA only occurred in samples of filtered water, but not in those containing only nematode genetic material. Sequencing and BLAST of the DNA fragments amplified by the MVC molecular marker, established a strong correlation of the amplified bands with proteins from betaproteobacteria species Overall, these results showed that the MVC marker is not related to a nematode virulence gene (avr) but to betaproteobacteria. New root-knot nematode resistant Mi-homologs were searched in accessions of the wild Solanum species. The S. huaylasense accession LA-1358 reduced reproduction of a population of M. arenaria to similar levels than the resistant tomato cultivar Anairis. Nevertheless, the resistance response of S. huaylasense accession LA-1358 was also nematode-species specific.

La regeneración de plantas a partir de explantes es la base de partida para poder aplicar tecnologías tales como la obtención de plantas haploides o la transformación genética. Este carácter presenta una amplia variabilidad inter e intraespecífica. Así, incluso dentro de la misma especie, podemos encontrarnos genotipos recalcitrantes cuya falta o escasa regeneración limita la aplicación de estas técnicas. Además del componente genético, otros factores que condicionan el éxito de la regeneración son: las condiciones fisiológicas del material de partida, los componentes del medio de cultivo, los reguladores de crecimiento, la temperatura, la luz, etc¿ La falta de información acerca de qué factores determinan que este proceso se produzca por una u otra vía morfogenética (la vía organogénica o la embriogénica) y la incertidumbre de cuantos genes están implicados, indica la necesidad de investigación básica de este proceso. El objetivo principal de este trabajo se centra en incrementar el conocimiento de la base genética así como la localización de QTLs implicados en la regeneración por la vía organogénica que es la predominante en tomate. Para ello, se han utilizado dos poblaciones de mapeo (F2, BC1) obtenidas por la Dra. Gisbert. La población F2 se obtuvo a partir de la autofecundación de una planta F1, resultante del cruce de una planta de tomate (S. lycopersicum L.) seleccionada por su escasa capacidad regenerativa (Anl27) y la accesión PE-47 de S. pennellii Correll. con alta capacidad de regeneración. Por su parte, la población BC1 se obtuvo a partir del cruce (Anl27 x F1). Con estos materiales se ha realizado una caracterización fenotípica y genotípica en clones de cada genotipo, que se han mantenido en cultivo in vitro. Utilizando el programa informático MapQTL¿ se han identificado seis QTL implicados en la regeneración localizados en los cromosomas 1, 3, 4, 7 y 8. Cinco de los QTLs (SpRg-1, Rg-3, SpRg-4a, SpRg- 4b, SpRg-7) proceden del tomate silvestre S. pennellii y uno (SlRg-8) procede de S. lycopersicum. El porcentaje de varianza explicada por cada QTL va desde el 7,4% al 27%, dentro del rango común (6-26%) registrado en el mapeo genético de QTLs relacionados con la regeneración in vitro en otros cultivos. SpRg-1 es el mayor responsable de la respuesta morfogenética mientras que SpRg-7 promueve el desarrollo del brote hacia una planta completa. Por otra parte los QTLs detectados en los cromosomas 8 (SlRg-8) y 4 (SpRg-4a, SpRg-4b) podrían contener genes que influyen en la formación de yemas y su desarrollo, respectivamente. Finalmente Rg-3, situado en la mitad del cromosoma 3 y ligado al gen de la invertasa ácida, se presenta como un alelo putativo del gen Rg detectado en S. peruvianum (Rg-1) y S. chilense (Dunal) Reiche. (Rg-2). Además del genotipo, el tipo y combinación de reguladores de crecimiento son importantes para el éxito de los protocolos de regeneración. En tomate, los reguladores de crecimiento más utilizados son las citoquininas, o la combinación de estas con auxinas. Otros reguladores como el etileno se han estudiado poco y los trabajos publicados muestran conclusiones dispares. Con el fin de estudiar la influencia del etileno en la regeneración se han realizado ensayos con dos compuestos liberadores de etileno (Ácido 1-aminociclopropano-1- carboxílico ¿ACC¿ y ácido 2-Cloroetil fosfónico ¿Ethephon¿) y dos inhibidores de etileno: AgNO3, que inhibe la acción del etileno; y CoCl2, que inhibe la producción de etileno. Los resultados obtenidos muestran que la concentración y el momento de la aplicación son dos factores fundamentales a tener en cuenta para el éxito de la respuesta organogénica. En concreto, la aplicación de inhibidores de etileno y su consecuente disminución tiene un efecto negativo sobre la regeneración en tomate ya que se disminuye y se retrasa la respuesta. Así mismo, las plantas regenerantes obtenidas presentan tamaño reducido, pudiendo aparecer vitrificación y malformaciones. Por otra parte, la suplementación de etileno puede mejorar la regeneración. Así, el número de plantas regeneradas a partir de explantes de S. pennellii se duplicó en los medios con ACC respecto al medio control. Si la aplicación de ACC se realiza tras la inducción de las yemas, pasados 10 días, el rendimiento total será mayor. Este resultado indica que éste compuesto podría ser utilizado para mejorar la regeneración en aquellos genotipos donde una vez formadas las yemas, el desarrollo de estas hacia plantas es el paso limitante. Por otra parte, las plantas regenerantes obtenidas muestran buen desarrollo, por lo que la suplementación de etileno no afecta al posterior crecimiento de las plantas regeneradas. Finalmente se ha estudiado la capacidad organogénica de las especies silvestres de tomate derivadas del complejo S. peruvianum L. sensu lato (s.l.): Solanum arcanum Peralta., Solanum huaylasense Peralta., Solanum corneliomulleri J. F. Macbr. y Solanum peruvianum L. sensu stricto (s.s.). Estas especies pueden tener un papel fundamental en la mejora, sobre todo de resistencias a factores bióticos. Sin embargo, debido a las barreras de hibridación que existen entre éstas y el tomate cultivado, no se han explotado en su totalidad en la mejora del cultivo. Para realizar cruces se ha recurrido a la fusión de protoplastos y el rescate de embriones. Sin embargo, no se ha estudiado la capacidad organogénica de las especies derivadas del complejo Peruvianum, lo que limita la aplicación de estas técnicas. En este trabajo se ha encontrado variabilidad intra e interespecífica en la capacidad organogénica de las especies silvestres derivadas de este complejo. En general, todas las accesiones ensayadas de S. corneliomulleri y S. huaylasense mostraron una elevada capacidad regenerativa y se han identificado accesiones recalcitrantes en S. arcanum (LA-2185) y S. peruvianum s.s (ECU-106 y CH-20). En este trabajo, también se ha realizado un análisis morfológico de las hojas que ha determinado que el número de foliolos y el nivel de dentición de los mismo pueden ser utilizados en la identificación in vitro de las especies del complejo. Sin embargo, el área de foliolo solo permite distinguir a S. arcanum del resto de especies del complejo. Finalmente se ha observado que las accesiones cuyas hojas tienen mayor número de foliolos y mayor nivel de dentición tienen una mayor regeneración.
Trujillo Moya, C. (2013). Estudio de la Capacidad Organogénica en tomate y especies relacionadas: Localización de QTLS implicados y estudio de la influencia del Etileno [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/31665
TESIS

A partir da identificação de catepsinas L em ensaios in vitro e em zimogramas partimos para purificação desta enzima no inseto. A região V2 foi selecionada como fonte de obtenção da cisteína proteinase já que dentre os três ventrículos o segundo apresentou maior atividade específica. Após diversas tentativas de isolar esta proteinase, foi estabelecida uma marcha de purificação que envolvia em todas as etapas a participação de metil metanosulfonato (MMTS), o que inativa a proteinase evitando assim autólise ao longo do processo de purificação. A marcha consistiu de três passos cromatográficos (troca-iônica, filtração em gel e afinidade, nesta ordem) onde foi observada a presença de duas cisteína proteinases, cada uma apresentando respectivamente as seguintes massas moleculares 32 e 45 kDa (SDSPAGE). As duas cisteína proteinases possuem o mesmo pH ótimo igual a 6,3. Além disso, estas enzimas foram termicamente inativadas a 40 ºC segundo uma cinética de primeira ordem aparente, sugerindo a existência de apenas uma espécie molecular de cada enzima na preparação com meia vida de 5 minutos para cis 1 e 4,8 minutos para cis 2. Foi determinada a constante de dissociação entre enzimainibidor, onde foi observado os valores de 17,3 nM para cis 1 e 7,11 nM para cis 2 através da titulação por E-64. A eficiência de catálise cis 1 e cis 2 é maior para o substrato sintético Z-FR-MCA do que para Z-RR-MCA, indicando que tratava-se de catepsinas L. Com o intuito de descrever os mecanismos moleculares por trás dos fenômenos fisiológicos no intestino médio do Hemiptera Dysdercus peruvianus foi construída uma biblioteca de cDNA a partir de mRNA deste tecido. Utilizamos ESTs provenientes desta biblioteca com o objetivo de identificar genes transcritos relacionados com proteínas de transporte de glicose além de enzimas digestivas. Após o processamento das leituras, surgiram 1053 ESTs úteis. Montando estes ESTs por alinhamento de bases, foram produzidos 62 contíguos e 841 singletos, o que totaliza 903 seqüências únicas. Entre as seqüências homólogas encontradas as mais relevantes para o nosso estudo foram: β-glicosidase (marcadora de membranas microvilares), α-glicosidase (marcadora de membranas 8 perimicrovilares), aminopeptidase (espaço perimicrovilar), catepsina L (conteúdo de vesículas secretoras) e proteína transportadora de açúcar do tipo GLUT. Estas seqüências encontradas tiveram a sua transcrição específica (ou preferencial) averiguada por RT-PCR semiquantitativo nos diferentes tecidos do inseto estudado (intestino médio, túbulo de Malpighi, corpo gorduroso, glândula salivar, ventrículo 1, ventrículo 2 e ventrículo 3 do intestino médio). O transporte de glicose e água in vivo foi estudado. Para isso, os insetos foram alimentados com uma solução contendo glicose e corante não absorvível seguido de dissecação e análise do conteúdo luminal. O transporte de água e glicose foi inibido por floretina 0,2 mM (inibidor do uniportador GLUT) e por florizina 0,1 mM (inibidor do simportador SGLT) e ativado por K2SO4 50 mM. Isto sugere a presença do transportador do tipo uniportador (GLUT), e do simportador K+-glicose (SGLT), ambos co-transportando água. O transcriptoma revelou proteína homóloga a transportador GLUT cuja seqüência está parcialmente completa e foi analisada com ferramentas de bioinformática
After identification of cathepsins L in vitro assays and in zimograms we began to purify this enzyme in insect midgut. The region V2 was selected as a source of material for purifying a cysteine proteinase because it contains most of the activity of that proteinase. After several attempts to purify this proteinase, an effective process was developed that avoid autolysis with methyl methanethiosulfonate (MMTS), a sulfhydryl-reactive and reversible sulfonating reagent for thiol-containing molecules. The purify process was made by three chromatographic steps (anion-exchange column, gel filtration column and affinity column in this order), where two cysteine proteinase were purified, cys 1 and cys 2 with 32 and 45 kDa (SDS-PAGE). The two cysteine proteinases have the same pH optimum of 6.3. Besides that, these enzymes were thermicaly inactivated following apparent first-order kinetics with a half-life of 5 min (cys1) and 4.8 min (cys2) at 40 ºC. Both Cys are inhibited by E-64 with a KD of 17.3 nM (Cys 1) and 7.11 nM (Cys2). Both Cys are more active on Z-FR-MCA than on Z-RR-MCA, suggesting they are cathepsins-L. With purpose of describe the molecular mechanisms underlying physiological phenomena in midgut of the Hemiptera Dysdercus peruvianus a cDNA library was prepared from midgut mRNA. We used ESTs from this library to identify transcripts genes related with glucose transport proteins besides digestive enzymes. Analysis of 1053 high-quality expressed sequence tags (ESTs) yielded 903 unique sequences comprised of 62 contigs and 841 singlets. Among the homologous sequences found the following are more relevant to our aim: β-glucosidase (microvillar membrane marker), α-glucosidase (perimicrovillar membrane marker), aminopeptidase (perimicrovillar space marker), cathepsin L (vesicles content) and sugar transporter protein, GLUT. These sequences had its specific transcription (or preferential) verified by semi-quantitative RT-PCR on different insect tissues (malpighian tubules, salivary gland, fat body, midgut, midgut first ventriculus, second ventriculus and third ventriculus). The glucose and water absorption across the first ventriculus of the midgut of the Hemiptera Dysdercus peruvianus were determined. The insects were fed with a 10 glucose-non-absorbable dye solution, followed by periodical dissection of insects and analysis of ventriculus contents. The transport of water and glucose can be inhibited by 0.2 mM phloretin (GLUT inhibitor) and by 0.1 mM phlorizin (SGLT inhibitor) and is activated by 50 mM K2SO4 The results suggest that D. peruvianus has a transporter uniporter like (GLUT) and K+-glucose symporter like SGLT, both co-transporting water. The transcriptome showed a GLUT homologous protein which sequence is almost complete and was analyzed by bioinformatics tools

Constitutionalizaton of Law’s different areas is a phenomenon gradually more ingrained in our cultural and legal framework. Maybe the best demonstration is the increasingly prominent role of the Constitutional Court (TC) – Constitution’s Supreme Interpreter – in defining and redefining concepts, rights and legal principles touching a range of subjects, from TaxLaw to Human Rights. This is relevant to understand the Law and its current effects whether it is valued positively or negatively.
La constitucionalización de las distintas áreas del Derecho es un fenómeno cada vez más arraigado en nuestra cultura y contexto jurídicos. Tal vez la forma más evidente en que se haya manifestado sea el creciente protagonismo del Tribunal Constitucional (TC), supremo intérprete de la Constitución, en la significación y resignificación de conceptos, derechos y principios jurídicos que, a través de sus sentencias, han afectado desde el derecho tributario hasta los derechos humanos. Sea que esto se valore positiva o negativamente, su relevancia para comprender el derecho y sus efectos en la actualidad es innegable.

Thesis (M.S. in Management)--Naval Postgraduate School, December 1990.
Thesis Advisor(s): Haga, William J. Second Reader: Gates, William R. "December 1990." Description based on title screen as viewed on March 29, 2010. DTIC Identifier(s): Life Cycle Costs, Cost Analysis, Weapon Systems, Military Procurement, Acquisition, Foreign Military Sales, USSR, Peru, Technology Transfer, International Trade, Debt, Theses. Author(s) subject terms: Peruvian Weapon Systems Acquisition Process, Life Cycle Cost Analysis, Unite Foreign Military Sales. Includes bibliographical references (p. 79-83). Also available in print.

El presente trabajo tiene como objetivo la elaboración del Plan de Negocios para la empresa “SAMI Peruvian Food & Drinks”, la misma que se dedica a la elaboración y distribución de comidas bajo el sistema del delivery. El objetivo de la organización es satisfacer la demanda de aquellas personas que buscan obtener un servicio que cumpla con todos los protocolos de bioseguridad establecidos, dada la pandemia a raíz de la llegada del COVID-19. Dichas personas mantienen la necesidad de compartir momentos agradables junto a familia, amigos y demás; sin embargo, la actual regulación en el país impide la atención de forma presencial, además del creciente temor social. Es por esto qué, se considera una excelente oportunidad de negocio el brindar gastronomía peruana dirigida a distintas personas y grupos con características y necesidades similares mediante el reparto a domicilio. El proyecto iniciará atendiendo operaciones en Miraflores, para lo cual se rentará el espacio con la capacidad instalada en cocina; únicamente para la elaboración de los pedidos. Dada la ubicación, se iniciarán las operaciones atendiendo en dicho distrito y los distritos límite: Barranco, Santiago de Surco, Surquillo y San Isidro. La recepción de los pedidos, así como la publicidad de la organización se manejará a través de medios digitales, tales como las redes sociales, puesto que éstas representan el menor costo posible para el inicio de las operaciones, además de ser en la actualidad un medio masivo de difusión y captación de clientes.
The goal of this work is to prepare the Business Plan for the “SAMI Peruvian Food & Drinks”, which is dedicated to preparing and distributing meals under the delivery system. The objective of the organization is to satisfy the demand of those people who seek to obtain a service that complies with all established biosafety protocols, given the pandemic following the arrival of COVID-19. These people maintain the need to share pleasant moments with family, friends, and others; however, the current regulation in the country prevents attention in person, in addition to the growing social fear. That is why, it is considered an excellent business opportunity to provide Peruvian food and drinks focused on different people and groups with similar characteristics and short-term needs through home delivery. The project will begin with operations in Miraflores, for which the space will be rented with the installed capacity in the kitchen, only for the preparation of orders. Given the location, operations will begin serving this district and the bordering districts: Barranco, Santiago de Surco, Surquillo and San Isidro. The reception of orders, as well as the advertising of the organization will be handled through digital sources, such as social networks, since these represent the lowest possible cost for the start of operations, in addition to being currently a means massive dissemination and recruitment of potential customers.
Trabajo de investigación

Peruvian Traditions is a small family business dedicated to the manufacture and export of alpaca garments. Currently, it has two brands White Label (Peruvian Traditions itself) and Norgäte and is aiming to expand its export process to Europe. Within the process of collecting information and analyzing the company, it was detected that the problem was the lack of differentiation between one brand and another which leads them to be "stuck in the middle". As a result, solutions were proposed for the problem determining that offering new product line called Alpaca Basics based on pre-existing garments within the Norgäte collection and on which it has been detected quite welcome, would be the best. The framework for this product line is the lean start-up methodology that is characterized by the formulation of a hypothesis and a testing process under continuous customer feedback that allows reaching a desired market-fit. Based on a previous analysis based on the segmentation, targeting and positioning (STP) the Netherlands was identified as target market. For the success of the product line, a 4P marketing campaign is required: product, place, price and promotion, which highlights the use and improvement of the current Peruvian Traditions website and a Google Ads campaign. To measure the results, three indicators will be used: number of visits through the web, conversion ratio and sales ratio. Subsequently, a scenario analysis is carried out taking into account three perspectives: negative, neutral and positive. Taking the neutral perspective as a reference, an increase in the number of visits is identified, from 1,460 to 4,380 visits per year. Which could generate an increase in the number of customers from 13 to 63.
Peruvian Traditions es una pequeña empresa familiar dedicada a la fabricación y exportación de prendas de alpaca. En la actualidad cuenta con dos marcas White Label (Peruvian Traditions) y Norgäte y se encuentra en la mira de expandir su proceso de exportación a Europa. Dentro del proceso de recopilación de información y análisis de la empresa se detectó que el problema era la falta de diferenciación entre una marca y otra lo cual los lleva a estar "stuck in the middle". A raíz de ello, se plantearon soluciones para la problemática, de ahí se determinó que la mejor solución sería lanzar una nueva línea de producto llamada Alpaca Basics basada en prendas preexistentes dentro de la colección de Norgäte. El marco para esta línea de producto es la metodología lean start-up que se caracteriza por la formulación de una hipótesis y un proceso de testeo bajo una continua retroalimentación del cliente que permite llegar a un anhelado market-fit. En base a un previo análisis, basado en la segmentación, focalización y posicionamiento (STP) se identificó que a Países Bajos como mercado objetivo. Para el éxito de la línea de producto se requiere de una campaña de marketing basada en las 4P: producto, plaza, precio y promoción, en donde destaca el uso y mejora de la actual web de Peruvian Traditions y una campaña de Google Ads que la respalda. Para medir los resultados se utilizaron tres indicadores: número de visitas a través de la web, ratio de conversión y ratio de ventas. Posteriormente se realiza un análisis de escenarios tomando en cuenta tres perspectivas: negativa, neutral y positiva. Tomando como referencia la perspectiva neutral, se identifica un incremento del número de visitas, pasando de 1,460 a 4,380 visitas por año.
Tesis

As sementes de Canavalia ensiformis são fonte de várias proteínas de interesse bioquímico e biotecnológico, como urease (JBU) e a canatoxina (CNTX). Ureases são enzimas dependentes de níquel que catalisam a hidrólise de uréia em amônia e carbamato encontradas em plantas, fungos e bactérias. A canatoxina, uma proteína tóxica isolada a partir da mesma semente em 1981 por Carlini & Guimarães é uma isoforma de urease. A CNTX e a urease possuem atividade inseticida, sendo que o efeito entomotóxico se deve à produção de fragmentos das proteínas, gerados pela ação das enzimas digestivas dos insetos. Apenas grupos de insetos com digestão baseada em enzimas acídicas são suscetíveis à toxina. O mecanismo de ação da CNTX e peptídeos gerados a partir da sua hidrólise não está claramente elucidado. Considerando o potencial das isoformas de ureases de C. ensiformis e peptídeos derivados para desenvolvimento de inseticidas, é importante investigar os mecanismos de ação e os órgãos ou tecidos–alvos de sua toxicidade nos insetos. Esse trabalho teve como objetivo testar diferentes protocolos visando a imunolocalização da toxina e/ou seus fragmentos em diferentes tecidos do inseto modelo Dysdercus peruvianus, um percevejo praga da cultura do algodão. Mesmo testando diferentes condições experimentais, observamos reação cruzada dos anticorpos anti-urease com os tecidos do inseto D. peruvianus, inviabilizando os estudos de imunolocalização. Considerando que a semente de algodão apresenta uma urease endógena ainda pouco caracterizada, e o alto grau de homologia existente entre ureases de diferentes fontes vegetais, a reação cruzada observada nos tecidos dos insetos pode ser devido à presença da urease do algodão e de seus fragmentos, provenientes do alimento consumido. Outras tentativas de identificação de alvos moleculares do inseto que interagem in vitro com a urease foram feitas, como ensaios de ligand blot e cromatografia de afinidade, mas estes não apresentaram resultados conclusivos. A produção de anticorpos monoclonais contra as diferentes isoformas de ureases de C. ensiformis está entre as alternativas possíveis para atingir nosso objetivo. Numa primeira tentativa, camundongos foram imunizados com uma mistura de isoformas da urease de C. ensiformis, que apresentam cerca de 85% de identidade, resultando na produção de anticorpos com especificidades diferenciadas para JBU e para a CNTX. Obtivemos 22 clones positivos: dois apresentaram maior especificidade pela CNTX (cerca de 4 vezes); dois clones foram mais específicos para urease (cerca de 2 vezes) e um foi capaz de reconhecer um peptídeo inseticida recombinante derivado da urease, com maior especificidade (resposta 1,5 maior do que CNTX ou JBU). Os resultados demonstraram que é possível produzir anticorpos monoclonais que diferenciam as diferentes ureases de C. ensiformis, para uso futuro em imunohistoquímica.

Ureases são enzimas que realizam hidrólise da uréia em amônia e dióxido de carbono e são isoladas de plantas, fungos e bactérias. A urease de C. ensiformis (JBU) tem algumas atividades biológicas independentes da sua atividade enzimática, como por exemplo: agregação plaquetária e efeito inseticida. O efeito inseticida é devido à liberação de um peptídeo interno por enzimas digestivas específicas dos insetos. Este peptídeo foi isolado, caracterizado e um análogo recombinante foi obtido e denominado Jaburetox-2Ec. Somente insetos com enzimas digestivas acídicas, como catepsinas, são capazes de liberar o peptídeo tóxico a partir da hidrólise de JBU. Insetos com enzimas básicas do tipo tripsina não são suscetíveis pois não há formação do peptídeo tóxico. Ninfas de D. peruvianus são sensíveis aos efeitos da JBU enquanto adultos não são. Este trabalho teve como objetivo estudar as diferenças enzimáticas entre os dois estágios do inseto para elucidar o processamento diferencial da JBU. Realizando a hidrólise in vitro da JBU com o homogeneizado de intestino de ninfas e adultos foi visto que tanto adultos como ninfas hidrolisam a JBU, mas somente ninfas liberam o peptídeo tóxico identificado pelo anticorpo específico anti-Jaburetox-2Ec. Ainda, através de ensaios enzimáticos utilizando substratos fluorogênicos e inibidores específicos, foi observada uma diferença no pH ótimo de atividade e na susceptibilidade a inibidores das enzimas digestivas presentes nos dois estágios. Substratos fluorogênicos correspondentes às regiões flanqueadoras do peptídeo dentro da JBU intacta foram desenhados e testados com os dois homogeneizados, o que também revelou que os homogeneizados dos dois estágios têm ação diferencial sobre estes substratos, sendo que ninfas liberariam mais eficientemente a extremidade Nterminal do peptídeo quando comparado aos adultos. Verificamos ainda que, em estudos eletrofisiológicos utilizando a técnica de “Planar Lipid Bilayer”, tanto a JBU como o Jaburetox-2Ec são capazes de se inserir em membrana lipídica planar, formando canais iônicos. Os canais formados pela JBU apresentaram quatro níveis de condutância majoritárias e seletividade para íons cloreto.
Ureases are enzymes that hydrolyze urea in ammonium and carbon dioxide and they have been isolated from plants, fungi and bacteria. Jackbean urease, from Canavalia ensiformis (JBU) displays biological activities unrelated from its enzymatic activity, as platelet aggregation and insecticide effect. This insecticide effect is due to the release of an internal peptide by insect specific digestive enzymes. This peptide was isolated, characterized and the recombinant peptide obtained was called Jaburetox- 2Ec. Only insects that rely on cathepsin-like digestive enzymes are able to hydrolyze JBU and release the toxic peptide. Insects with alkaline enzymes like tripsins are not susceptible because they don’t release the toxic peptide. Nymphs of D. peruvianus are susceptible to JBU effects while adults are not. The goal of this work was to study the enzymatic differences between both insect stages to elucidate JBU’s differential processing. In vitro hydrolysis were performed with nymphs and adults midgut homogenates and we observed that both adults and nymphs hydrolyze JBU but only nymphs are able to release the toxic peptide identified by Jaburetox-2Ec antibodies. Furthermore, in enzymatic assays using different fluorogenic substrates and specific inhibitors a difference in optimum pH and susceptibility to inhibitors of the digestive enzymes in both stages was observed. Fluorogenic substrates corresponding to the flanking regions of the peptide inside the intact JBU were produced and tested with both homogenates. Homogenates from both stages have differential action upon these substrates, considering that nymphs hydrolyses more efficiently the N-terminal of the peptide compared to adults. In electrophysiological studies using the Planar Lipid Bilayer technique we verify that JBU and Jaburetox-2Ec are able to insert in planar lipid membrane forming ionic channels. JBU’s channels display four major conductance levels and selectivity for chloride íons.

El incremento de la migración de ciudadanos peruanos a Buenos Aires constituye un fenómeno reciente, y la intensificación del desarrollo organizacional y de la acción pública de los mismos encuentra su correlato en la participación en fiestas, organizaciones civiles, entre otros. Este artículo es un avance de investigación y se centra en los grupos de danzas peruanas en Buenos Aires con el propósito de describir algunas de las modalidades de enseñanza de la danza peruana. Así, veremos cómo las interacciones intergrupales y de los grupos de danza con el Estado peruano en Buenos Aires, la necesidad de «mostrar el hacer» así como las formas de participación en los grupos y los aprendizajes previos de las danzas con los que se insertan los bailarines en estos ámbitos, configuran procesos de educación no formal. El análisis se basa en una dimensión microanalítica que refiere a las representaciones y prácticas de bailarines y directores de grupos. El trabajo de campo se focalizó en el período 2005-2008, constituyendo las unidades de estudio cuatro grupos de danzas folklóricas peruanas. Finalmente, se muestra que estos espacios poseen un potencial intercultural para trascender las manifestaciones culturales, promoviendo trabajos de concientización que incluyan a un público no peruano.
The increasing migration of Peruvian citizens to Buenos Aires is a recent phenomenon, and the intensification of the development of organizations and their public presence are correlated with the participation in festivities, civic organizations and the like. This article is a preliminary report on research that focuses on Peruvian dance groups in Buenos Aires and that proposes to describe some of the ways that Peruvian folkdances are transmitted. I show how a process of non formal education is shaped by the interactions among dance groups and between them and representatives of the Peruvian Government in Buenos Aires; and how it is influenced by the need to «demonstrate through performance,» by ways of participating in the groups and by the previous knowledge of the dances that the performers bring with them. The analysis focuses on the microanalytic dimension of representations and practices of the dancers and directors of the dance groups. The fieldwork was carried out in the period 2005-2008 and focused on four groups of Peruvian folk dancers. The phenomena studied are shown to have great potential as intercultural mediators that transcend cultural manifestations to become mechanisms of consciousness raising that reach out to publics beyond the Peruvians themselves.

Thesis (M.A.)--University of California, San Diego, 2008.
Title from first page of PDF file (viewed July 22, 2008). Available via ProQuest Digital Dissertations. Includes bibliographical references (p. 120-125).

This study stressed the Rorschach-Content area in 237 Lima inhabitants. The Contents are an indicator that helps to understand the basic characteristics of personality (self-perception,interpersonal relations, cognitive mediation and ideation). This descriptive study used an accidental-probabilistic sample. The variables were age, gender, schooling, The descriptive statistic was used to analyze demographic contents and variables, and the non-parametric statistic (Kruskall-Wallis) to compare data. The results stresses the absence of religious contents (R 1) and the importance of anatomic contents (An) across the variables. The variable gender shows differences: men are interested in culture and show achievement motivation. Women are interested in home. With regarded lo schooling, the group including superior educated and complete secondary educated students shows solidarity signs, interest in socialization and ahigh cognitive level.
Se investigó el área de Contenidos Rorschach en 237 habitantes de Lima. Los Contenidos constituyen un indicador para la comprensión de características básicas de la personalidad (auto-percepción, relaciones interpersonales, mediación cognitiva e ideación). El estudio fue descriptivo con muestreo probabilístico accidental. Las variables fueron edad, género, nivel de escolaridad. Se utilizó estadística descriptiva para analizar contenidos y variables demográficas, y no paramétrica (Kruskall-Wallis) para comparar datos. Los resultados destacan la ausencia de contenidos religiosos (Rl) y la importancia de contenidos anatómicos (An) entodas las variables. Los géneros difieren significativamente: los hombres interesados en lacultura y afán de logros y las mujeres, en el hogar. En cuanto a nivel de escolaridad, el grupo de educación superior y secundaria completa obtiene signos de solidaridad, interés en la socialización y mayor riqueza cognitiva.

Los programas de tesis digitales en Perú se iniciaron el 2002, Cybertesis fue la primera plataforma adoptada, actualmente DSpace es el software de uso oficial. En Junio de 2013 se publicó la Ley 30035 que obliga a las entidades públicas implementar un repositorio digital de acceso abierto, así mismo, crea el Portal “ALICIA”. En Setiembre de 2016 entró en funcionamiento el Portal “RENATI”, plataforma que difunde las tesis de entidades peruanas de educación superior. Actualmente ALICIA tiene 194 repositorios y 271,000 publicaciones en acceso abierto. Por su parte RENATI tiene 170 repositorios de tesis y 218,397 documentos de acceso abierto y restringido. Las políticas institucionales y normativas de gobierno han promovido un crecimiento acelerado de los repositorios, en especial de las tesis, de igual forma, han generado nuevos servicios en la mejora de la calidad de las tesis.
The Peruvian ETD programs began in 2002, Cybertesis was the first platform adopted and currently DSpace is the official software. In June 2013, Law 30035 was published that forces public institutions to implement an open access digital repository, and creates the “ALICIA” Portal. In September 2016, Portal “RENATI” was published, it´s a platform that disseminates the Peruvian digital thesis. “ALICIA” currently has 194 repositories and 271,000 open access publications. RENATI includes 170 digital thesis repositories and 218,397 open and restricted access digital theses. Government and regulatory policies has promoted an accelerated growth of repositories, especially digital theses; in the same way, new services to improve the quality of the thesis were developed.

Antimicrobial resistance and antimicrobial misuse are of global concern but affect more the resource-constrained settings. Limited research has been done in Peru in order to address these topics. We interviewed 256 physicians of two hospitals of Lima in order to assess the knowledge, attitudes, and practices about antimicrobial resistance and antimicrobials use. Most of them agreed that antimicrobial resistance is a problem worldwide and within Peru, but only 20% correctly estimated the level of resistance of Klebsiella pneumoniae to third generation cephalosporins. We also developed a network of nine hospitals in Lima to perform a resistance surveillance of key isolates causing bacteremia. We found during a one-year period that Staphylococcus aureus was the most frequently Gram positive isolated (22%). K. pneumoniae and Escherichia coli were the most frequent Gram-negative bacilli, >75% of both produced extended-spectrum β-lactamases (ESBLs). ESBL-producers microorganisms had also higher level of co-resistance to ciprofloxacin and gentamicin. Finally, to explore the antimicrobial resistance and molecular characteristics of methicillin-resistant S. aureus, we analyzed isolates from patients and healthcare workers (HCWs). Fifty percent of 338 blood isolates were methicillin-resistant (MRSA); one predominant multidrug-resistant MRSA strain was found in all hospitals (ST 5 spa t149-SCCmec I-, the Cordobes-Chilean clone. MRSA nasal carriage rate of 8.7%, was found among HCWs. The two most common clones circulating among HCWs were also the two predominant among patients with bacteremia. There is a need to implement cost-effective infection control policies to reduce the transmission of multidrug resistant microorganisms in these settings.
La résistance aux antibiotiques et leur utilisation inappropriée constituent un problème majeur de santé publique qui affecte particulièrement les pays à revenus faibles et intermédiaires. Les données concernant cette problématique sont très limitées au Pérou. Nous avons interrogé 256 médecins de deux hôpitaux à Lima afin d’évaluer leurs connaissances et leurs pratiques vis-à-vis de l’utilisation des antibiotiques et de la résistance aux antibiotiques. La plupart des médecins étaient conscientisés à la problématique de la multirésistance chez les entérobactéries, mais seulement 20% d’entre eux estimaient correctement le taux résistance de Klebsiella pneumoniae aux céphalosporines à large spectre au sein de leurs institutions. Au cours de ce travail, un réseau entre neuf hôpitaux de Lima a été implémenté afin de suivre la résistance aux antimicrobiens des souches isolées d’hémocultures. Au cours des années 2008-2009, le Staphylococcus aureus était la bactérie Gram-positive la plus fréquente (22%) ;K. pneumoniae et Escherichia coli représentaient les bactéries Gram-négatives les plus souvent isolées d’hémocultures. Parmi ces dernières entérobactéries, 75% produisaient des β-lactamases à spectre étendu (BLSE) avec un taux élevé de co- résistance à la ciprofloxacine et à la gentamicine. Les souches de S. aureus isolées d’hémocultures chez les patients hospitalisés et de dépistage chez les professionnels de santé (PS) ont été caractérisées pour leur profil de résistance aux antibiotiques et génotypées par biologie moléculaire. La moitié des 338 souches isolées d’hémocultures étaient résistantes à la méticilline (« MRSA ») et appartenaient à un clone prédominant disséminés dans ces différents hôpitaux, appelé le ST 5- spa t149-SCCmec I, ou le clone Cordobes-Chilien. Le taux de portage nasal de MRSA parmi les PS était à 8,7% ;les deux génotypes les plus fréquemment observés chez les PS appartenaient aux mêmes clones que ceux retrouvés majoritairement chez les patients hospitalisés. Cette observation suggère une transmission horizontale. L’implémentation de politiques de contrôle de l'infection est primordiale dans les établissements de santé au Pérou afin de réduire la transmission de micro-organismes multi-résistants.
Doctorat en Sciences biomédicales et pharmaceutiques (Médecine)
info:eu-repo/semantics/nonPublished

A cultura algodoeira tem significativa participação na economia mundial. O algodão (Gossypium hirsutum) é predado por uma grande variedade de insetos, entre eles o percevejo manchador Dysdercus peruvianus (Hemiptera: Pyrrhocoridae). O hábito alimentar deste inseto provoca danos nas sementes e fibras depreciando-as para a utilização comercial. Como alternativa para o aumento da resistência das plantas ao ataque de insetos predadores, estratégias biotecnológicas vêm sendo consideradas. A Canavalia ensiformis é fonte de proteínas com atividade inseticida como a urease, a canatoxina e a concanavalina-A. Tanto a canatoxina como a urease exercem efeitos tóxicos em ninfas deste percevejo, causando mortalidade e atraso no desenvolvimento dos insetos sobreviventes. De maneira análoga a outros modelos de insetos já estudados, a toxicidade em D. peruvianus dependeria da ativação proteolítica das proteínas por enzimas digestivas (catepsinas) do inseto e formação de um peptídeo tóxico. Neste trabalho, estudos de hidrólise in vitro na presença de inibidores seguidos por SDS-page e Western-blot confirmaram a ativação proteolítica da urease por enzimas do tipo catepsina-B. O homogeneizado de intestinos de ninfas hidrolisa também substratos típicos para catepsinas, como hemoglobina e Abz-AIAFFSRQ-EDDnp, e a atividade proteolítica é bloqueada por pepstatina-A e E-64, inibidores específicos para aspártico e cisteíno proteinases, respectivamente. Duas cisteíno proteinases (DpQ-NR e DpQ-E) de intestinos de ninfas de D. peruvianus foram parcialmente purificadas e caracterizadas. Ambas apresentaram pH ótimo de atividade proteolítica entre 4,5 e 6,5, na presença de DTT e foram totalmente inibidas por E-64. No entanto o perfil de inibição evidencia a presença de outras enzimas nas frações. Ensaios de especificidade da fração DpQ-NR revelaram a preferência da enzima por substratos contendo resíduo hidrofóbico em P2, enquanto a enzima DpQ-E tem preferência por substratos contendo aminoácidos básicos em P1. Assim, o trabalho desenvolvido sugere a presença de proteinases múltiplas no sistema digestivo de D. peruvianus e o possível envolvimento destas peptidases na ativação proteolítica da urease.
Cotton (Gossypium hirsutum), an important agricultural commodity, is attacked by a number of pests, such as the cotton stainer bug Dysdercus peruvianus (Hemiptera: Pyrrhocoridae) which may cause severe losses in cotton plantations. Bug feeding on the cotton seeds and stains the cotton fibers resulting in damage to the seed, besides being a vector for phytopathogenic bacteria and fungi. Biotechnology strategies focusing on plant defensive molecules are being considered as an alternative to increase host plant resistance against this pest. The jack bean Canavalia ensiformis is the source of proteins displaying insecticidal activity such as urease, canatoxin and concanavalin-A. Canatoxin and urease were shown to cause lethality and severe detrimental effects in surviving insects, delaying the development stages of D. peruvianus. As in other insect models studied previously, this would depend on the proteolytic activation of the protein by insect cathepsin-like digestive enzymes to produced toxic peptides. The in vitro hydrolysis of urease in the presence of inhibitors, followed by SDS-page and Wester-blot confirm the proteolytic activation by insect cathepsin B-like enzymes. A nymphal gut extract hydrolysed typical cathepsin substrates, such as hemoglobin and Abz-AIAFFSRQ-EDDnp, and hydrolysis was blocked by pepstatin-A and E-64, specific inhibitors of aspartic and cysteine proteinases. Two cysteine peptidases (DpQ-NR and DpQ-E) of D. peruvianus nymphs were partially purified and characterized. Both enzymes showed maximum activities at pH 4.5-5.0 and 5.5-6.5 in the presence of dithiotreitol and were totally inhibited by E-64. However, the inhibition profile points out to the presence of other types of proteinases in both fractions. DpQ-NR preferentially hydrolysed substrate with hydrophobic amino acids in P2, while DpQ-E had preference for substrate with basic residues in P1. Therefore, our work suggests the presence of multiple proteinases with distinct biochemical properties in the digestive system of D. peruvianus and the potencial involvement of these peptidases on the proteolytic activation of urease.

O controle biológico pode ser definido como a utilização de antagonistas naturais para a diminuição ou redução de determinado organismo indesejado. Atualmente, esta estratégia constitui uma ferramenta segura e efetiva tanto para o controle de pragas da agricultura e da pecuária, quanto para vetores de doenças que atingem a saúde pública. O fungo filamentoso Metarhizium anisopliae tem se destacado neste âmbito devido às suas características entomopatogênicas e acaricidas, sendo utilizado comercialmente no Brasil para o controle da cigarrinha da cana-de-açúcar e estudado, mundialmente, como potencial biopesticida. Não se sabe quais são os fatores que definem a predileção do fungo a determinados hospedeiros, mas se acredita que fatores tais como uma maior adaptação do patógeno, a composição e a morfologia da cutícula possam influenciar. Neste trabalho foi avaliada a capacidade entomopatogênica de 8 linhagens de M. anisopliae sobre dois artrópodes: o percevejo manchador do algodão Dysdercus peruvianus e a lagarta da soja Anticarsia gemmatalis. Todas as linhagens testadas de M. anisopliae foram mais efetivas contra D. peruvianus, quando comparadas à A. gemmatalis. A linhagem CG47 mostrou-se eficaz contra ambos os artrópodes testados. As linhagens C12, CG47, CG97 e Nordeste foram ativas contra A. gemmatalis, sendo C12 a mais virulenta. A secreção de proteases e quitinases durante o cultivo do fungo, em presença de cutículas de artrópodes, foi avaliada e parece ter sido influenciada pelo tipo de fonte de carbono disponível no meio de cultivo. A secreção de quitinases, ao contrário das proteases, pareceu não sofrer repressão tão acentuada, com a utilização de fontes facilmente metabolizáveis como a glicose; e, embora existam grandes diferenças entre as linhagens na capacidade de secreção de ambas as enzimas, a virulência observada nos bioensaios pareceu não estar correlacionada com a capacidade quitinolítica ou proteolítica das diferentes linhagens. Levando-se em conta que o processo de infecção de M. anisopliae em hospedeiros é multifatorial e dependente também dos artrópodes-alvo, foi feita a contagem total de hemócitos, sendo possível verificar um aumento destas células de defesa logo após o desafio com o patógeno. Neste trabalho, fica evidente o potencial de determinadas linhagens de M. anisopliae para o controle de D. peruvianus e A. gemmatalis. Também é sugerida a utilização do percevejo manchador do algodão como um novo modelo de pesquisa para os estudos que envolvem as interações patógeno-hospedeiro, mediante a análise de novos fatores de virulência desse patógeno e da resposta do hospedeiro à infecção.
Biological control can be defined as the utilization of natural enemies to reduce or to diminish some unwanted organism. Nowadays, this strategy is a safe and effective tool to control plagues from agriculture and pecuary and also for vectors of some important diseases of public health. The filamentous fungi Metarhizium anisopliae have been standed out in this field because its entomopathogenic and acaricide characteristics, being commercially used in Brazil to control the sugar cane bugs and being studied worldwide as a potential biopesticide. The factors that define the fungi specificity for some hosts are not known, although, is believed that factors as a greater adaptation of the pathogen and the cuticle composition and morphology may influence. In this work the entomopathogenic activity of 8 strains of M. anisopliae was evaluated against two arthropods: the cotton stainer Dysdercus peruvianus and the velvet bean caterpillar Anticarsia gemmatalis. All M. anisopliae strains tested were more efficient against D. peruvianus when compared to A. gemmatalis. Strain CG47 was active for both arthropods tested. Strains C12, CG47, CG97 and Nordeste were active against A. gemmatalis and C12 was the most virulen strain. Protease and chitiunase secretion during fungi growth with arthropod cuticles was tested and seem to be influenced by the kind of carbon source available in the media. Chitinase secretion, on the contrary to proteases seem not to respond so effectively to repression by the utilization of simple carbon compounds as glucose and, despite there are big differences between strains in the capacity to secret both enzymes, the virulence observed during bioassays seemed not to be correlacionated to their proteolitic or chitinolitic activity. Aware that the M. anisopliae infection process on hosts is multifactorial and also depends on arthropod response, the total hemocytes count were made and was possible to detect an increase on cellular counts after challenge with the pathogen. In this work is evident the potential of some M. anisopliae strains to control D. peruvianus and A. gemmatalis and it also suggests the utilization of the cotton stainer as a model for researches about host-pathogen interactions, studies about pathogen virulence factors as well studies to elucidate the host response to infection.

This    qualitative research investigates the social positions of Peruvian female migrants in the Catalan labour market in Spain. It questions how social categories interactin order to determine the social positions of individuals, a nd how the positions can be related to a global  world structure.   Four unstructured life story interviews with  Peruvian women resident in  Catalonia were realized in December 2014. The interviews and two previously made studies have then been reviewed by a narrative analysis. The research formed a theoretical framework of intersectionality interpreted by Nina Yuval-Davis, and a globalization and female migration theory by Saskia Sassen.  The analysis with the  implementation of theories showed that nationality has a particularly strong influence in the intersection of social categories, though one must understand how all the identifications lay imbedded in each other in order to determine the social position of each individual. It was  also concluded that an intersection of the identifications of the individuals interacts with global structures in order to determine the  social positions of the study participants in the Catalan labour market.The city of Barcelona  could through the lives of the four Peruvian females be seen as an economic centre that demands cheap labour by migrants, and where the social positions in the labour market can show a division of core and peripheral countries.

Os insetos da ordem Hemiptera apresentam membranas lipoproteicas que revestem as microvilosidades das células intestinais, como se fossem dedos de luva, e formam expansões para o lúmen do intestino que parecem terminar em fundo cego. A presença das duas membranas no ápice dos enterócitos gera questões intrigantes de como se dá a formação da membrana perimicrovilar, como ocorre a absorção de nutrientes e como se dá o encaminhamento de enzimas digestivas para o lúmen intestinal. A digestão de proteínas baseada em enzimas originalmente lisossômicas é uma característica marcante nos Hemiptera, em especial os Heteroptera que evolutivamente voltaram a uma alimentação de polímeros. O presente estudo indica que os genes das proteinases tipicamente lisossômicas sofrem uma série de duplicações, havendo a manutenção de um gene para função puramente lisossômica e a divergência funcional dos demais genes para a função de digestão extracelular. O gene que mantém a função lisossômica não é modulado pela alimentação, além de ser expresso nos mais diversos tecidos. Já os genes que se especializaram na digestão extracelular têm a expressão aumentada com a ingestão de alimento, indicando sua função. Parece não haver diferença nas características relacionadas ao encaminhamento celular das proteínas produzidas por esses genes, indicando que o direcionamento para a rota secretória é devido à superexpressão dos genes relacionados à digestão. Enzimas como alfa-glicosidases, alfa-manosidases e aminopeptidases que participam da digestão extracelular seguem a rota secretória que envolve a formação de vesículas de dupla membrana. Foi possível ampliar o modelo de digestão incluindo a participação das catepsinas D, de uma alfa-glicosidase solúvel e a possível participação de uma tiolredutase além do definir o local de atuação das lipases. Temos agora uma visão global da participação das enzimas digestivas que atuam na digestão em D. peruvianus.
The insects of the order Hemiptera have lipoprotein membranes lining the microvilli of midgut cells, like glove fingers, and form expansions into the lumen of the intestine. The presence of two membranes on the apex of enterocytes thus generates intriguing questions about the formation of perimicrovillar membrane, the absorption of nutrients, and the targeting of digestive enzymes into the intestinal lumen. The digestion of proteins based on originally lysosomal enzymes is an important feature in Hemiptera, especially in Heteroptera that evolutionary returned to feed on polymers. The genes of typical lysosomal proteinases undergo a series of duplications followed by the maintenance of a gene for purely lysosomal function and functional divergence of other genes for extracellular digestion function. The gene that maintains the lysosomal function is not modulated by feeding, in addition to being expressed in diverse tissues. By the other hand, genes specialized in extracellular digestion are up regulated by food intake, indicating its function. No difference was found in the targeting in the proteins produced by these genes, which indicates that targeting to the secretory route is due to overexpression of digestion-related genes. Enzymes involved in extracellular digestion as alpha-glucosidase, alpha-mannosidase and aminopeptidase follow the secretory route that includes the formation of double membrane vesicles. In this study we increased the digestion model adding the participation of cathepsin D, a soluble alpha-glucosidase, and the possible participation of a tiolredutase, and also to defining the place of lipases operation. We now have a global view of the participation of digestive enzymes involved in digestion D. peruvianus.

This study examines whether aquaculture has the potential to reduce deforestation in the Peruvian Amazon. The natural resources of the Peruvian Amazon are subject to extreme pressures due to increases in subsistence farming, cattle ranching, and logging in the region. The resulting loss of biodiversity has affected the delicate soil balance that is characteristic of the Amazon, and has contributed to water pollution as well as erosion (Guerra et al. 2001). One of the highest rates of deforestation in the Amazon basin can be found at the foothills of the eastern Andes (Lepers et al. 2005), which includes the area in this study, located in the Peruvian state of Amazonas. In this part of the Amazon, deforestation is mainly caused by small-scale subsistence agriculture (Achard et al. 1998) such as that found in the communities of Condorcanqui. Fishing is an essential part of the socio-economic system that functions in the Peruvian Amazon. Fish meat is the most important source of animal protein in the Amazon, and the main generator of cash for indigenous people (McDaniel, 1997). However, freshwater Amazonian fisheries have been subject to extreme overexploitation in the past few decades (Rainforest Conservation Fund, 1999). Boats with technological equipment and large-scale capacities have threatened stocks in local rivers and oxbow lakes, which has affected the ability of small-scale, native fisherman to support themselves (Rainforest Conservation Fund, 1999). The Instituto de Investigaciones de la Amazonia Peruana (IIAP) has assisted the development of fish farming in the Condorcanqui region as a way to augment diets and decrease the ecological impact of subsistence farming. Fish farming can also be seen as a way to substitute for the loss of traditional river fishing. IIAP has become the leading governmental organization in fisheries research and aquaculture in the Peruvian Amazon, and works to provide native-species fingerlings, and educational courses to native aquaculturists. This study surveys the subsistence villages along road and river communities to determine the impact of fish farming on deforestation in the Condorcanqui region. This region is populated by small communities of indigenous Awajún and Wampí tribesmen, who practice subsistence agriculture. Data was collected from a sample of 184 families in ten different communities. Five of the villages were situated along the banks of the Nieva or Santiago river systems, while the other five were accessible by road, travelling southwest from the town of Santa Maria de Nieva. Data was collected with the assistance of the Research Institute of the Peruvian Amazon (IIAP), which provided a guide who had relations with all of the communities. All of the families in the sample practice subsistence agriculture, while 104 of the respondents supplement their agricultural crops with fish from aquaculture ponds integrated into their farmland. The participants answered a range of questions about the size of their farms, and the productivity of their land. We use a variety of regression-based approaches to determine how incorporating aquaculture into subsistence farmlands affects deforestation rates after controlling for socioeconomic and farm characteristics. Our study suggests that an extra square meter of aquaculture reduces the area deforested for crops on approximately a one for one basis. However, aquaculture should maintain its productivity for much longer than cropping, as it does not depend on soils whose fertility can be exhausted in a few years. Our simulations, based on our survey results, indicate that over time aquaculture should reduce deforestation significantly, especially in areas where soils provide only a few years of subsistence crops. These should be regarded as interesting but preliminary results. Because we used a convenience-based sampling approach, our results could be affected by selection bias. In addition, we do not have enough information to test whether selection bias in the implementation of fish farming affect our results. Therefore, these results suggest that aquaculture could be useful in limiting deforestation, but additional work should use experimental methods or more in-depth surveys to measure the effect of aquaculture on deforestation.

Thesis (M.S. in Defense Analysis)--Naval Postgraduate School, December 2005.
Thesis Advisor(s): Gordon McCormick, George Lober. Includes bibliographical references (p. 55-58). Also available online.

This thesis is a study of the ways in which the concept of sexuality changes in meaning over time and as it travels through different Peruvian feminist discursive fields and sites of action. Although academic research on Latin American feminists' politics of sexuality has been very limited to date, their own documents reveal a notable absence of debates on same-sex sexuality within the field of sexual and reproductive rights. Moreover, when same-sex sexuality is incorporated into discourse, conceptualized as lesbian rights, it does not refer to low-income women. This paradox is mirrored in feminist connections with British international development agencies. Combining multisited ethnography with a variety of qualitative research methods, I examine the effects of the shifting meanings and normative assumptions in the deployment of 'sexuality' by key Peruvian feminist individuals and organizations in their work at local, national, regional and global levels. In this thesis I show that feminist discourse on sexual rights is implicitly heteronormative with reference to women in Lima's low-income settlements and related national policy advocacy. This limitation is influenced by and reinforces the discursive and funding pressures emanating from international development agencies. Meanwhile, the same feminist actors' national and regional public policy advocacy on lesbian rights and non-discrimination on the basis of sexual orientation does not connect sexuality with, class, ethnic or national differences and inequalities. The consequence is the construction of a normative middle class, modern, global, urban lesbian subject in Peru that is considered of little relevance by UK-based international development agencies. At regional and global levels, feminist discourse on sexual and reproductive rights has very recently incorporated ythe notions of 'sexual and gender diversity', thus incorporating questions of same-sex sexuality. However, differences of social class, race, ethnicity and geo-political location remain unexamined. I argue that in future feminist cultural politics, the pre-theoretical commitments and normative assumptions of sexuality and related rights-based concepts, deployed in different discursive fields and sites of action, need be theorized on the basis of women's knowledge from different economic, ethnic, racial and national contexts.

The author proposes the text for a gender identity law based on the parameters of conventionality of the Inter-American Court of Human Rights and experiences of comparative law on the basis of depathologization trans identities.
La autora propone el texto para una ley de identidad de género basado en los parámetros de convencionalidad de la Corte Interamericana de Derechos Humanos y las experiencias del derecho comparado sobre la base de la despatologización de las identidades trans.

First, this work proposes a study of the origin, elements and application of the Compliance Program or, also named, Regulatory Compliance Program. The aforementioned program is understood as an internal device that corporations use not only to comply with the current legislation but also to prevent and detect legal violations they could be found in or as part of the activities they carry out.Second, it tries to explain and develop the connection between, on one hand, the new risks in the financial and technological development and, on the other hand, Criminal Law as a protective body of important legal assets for society. The aim is to analyze Criminal Compliance to get the corporationto manage its activities under current legislation, especially Criminal Law.In this regard, it enlarges the sectoral developments based on the Peruvian Government’s implementation of the the Compliance Program in the legal framework.Finally, it analyzes the impact of Criminal Compliance in the criminal legal framework. For that end, it refers to the criminal liability system in Peru and in what way it impacts on the application of Criminal Compliance.
El trabajo propone, en primer lugar, un estudio del origen, los elementos y la aplicación del Compliance Program o, también llamado, Programa de Cumplimiento Normativo, entendido como un dispositivo interno que las empresas implementan para cumplir con la normatividad vigente, así como para prevenir y detectar las infracciones legales que se produzcan dentro de las mismas o como parte de las actividades que estas realizan. Asimismo, se intenta explicar y desarrollar la relación entre los nuevos riesgos, debido al desarrollo económico y tecnológico, y el derecho penal como ente protector de bienes jurídicos de importancia para la sociedad. Esto último tiene como finalidad analizar el Criminal Compliance, destinado a que la empresa ordene su actividad conforme a la normativa aplicable, en especial la ley penal. En tal sentido, se desarrolla la aplicación que el Estado peruano ha realizado del referido Programa de Cumplimiento Normativo en el ordenamiento jurídico, en concreto los avances sectoriales. Finalmente, se analiza el impacto del Criminal Compliance en el ordenamiento jurídico penal. Para ello, se hace una referencia al sistema de responsabilidad penal adoptado en el Perú y de qué manera esto impacta en la aplicación del Criminal Compliance.