La Llama es el Camélido doméstico más abundante de Argentina. La cría de Llamas constituye una actividad económica de gran importancia debido a que es una especie poliproductora de carne, fibra, cuero y transporte. Actualmente existe interés creciente por mejorar el rendimiento y calidad de estos productos. El estudio de genes candidatos permite vincular las variaciones de un carácter y la manera en que éste se manifiesta en el fenotipo del individuo. En ese contexto la hormona de crecimiento (GH), producto del gen GH1 y secretada por la glándula pituitaria, estimula el crecimiento de huesos y músculos. Por otra parte, es bien conocido el interés que existe en la producción de fibras por determinados colores con mayor valor comercial. La determinación del color de capa en mamíferos se debe a la interacción de los genes MC1R (receptor 1 de melanocortina) y ASIP (péptido de señalización Agouti). Ambos controlan el tipo y localización de pigmento eumelánico (negro-marrón oscuro o sepia) o feomelánico (rojoamarillento) producido. Esta Tesis tuvo por objetivo caracterizar y analizar la diversidad genética del gen GH1 y realizar la caracterización molecular de las variantes alélicas de MC1R y ASIP en Llamas con distintos fenotipos de colores de capas. Mediante PCR se amplificaron y luego secuenciaron los tres genes. El gen GH1 mostró un alto nivel de variabilidad encontrándose 15 SNPs, mayormente situados en región no codificante. Sin embargo se identificaron dos polimorfismos en el promotor y uno en la región 5´no traducible. Dado que los polimorfismos localizados en el promotor podrían afectar los niveles de expresión del gen, se concluye que los mismos pueden ser útiles en futuros estudios de asociación. Con respecto al gen MC1R se identificaron 13 SNPs en región codificante, 10 de los cuales fueron no sinónimos. La combinación de 3 de estos polimorfismos permitieron diferenciar Llamas con capas pigmentadas (A259/A376/T383) de las blancas no albinas (BNA) que carecían de pigmento (G259/G376/C383). En ASIP el hallazgo más importante fue una deleción de 57pb en el Exón 4 con posible pérdida de función. De esta manera, el alelo delecionado en homocigosis se observó en Llamas eumelánicas y el alelo sin delecionar en estado homocigota o heterocigota, se vió en Llamas feomelánicas. La identificación de los alelos de los genes MC1R y ASIP permitió proponer un mecanismo por el cual se genera la pigmentación feomelánica y eumelánica (TO) en las Llamas.

The myc family of oncogenic transcription factors, which includes c-myc, N-myc and L-myc, control major cellular processes such as proliferation and differentiation by integrating upstream signals and orchestrating global gene transcription. They do this largely through dimerising with Max, which together bind to enhancer (E)-box elements in DNA. Myc proteins function similarly but differ in potency and tissue distribution. For instance, N-myc is expressed predominantly during development in undifferentiated cells of the nervous system, whereas c-myc is ubiquitously expressed in all proliferating cells. Myc proteins, when deregulated, are major drivers of tumourigenesis. Myc deregulation occurs in up to 70% of all human cancers and is often associated with the most aggressive forms. For example, MYCN, the gene encoding N-myc, is amplified in 20-30% of neuroblastomas, and amplification strongly correlates with advanced stage and poor prognosis. Myc proteins are therefore considered “most wanted” targets for cancer therapy, but have long been considered undruggable mainly due to challenges in nuclear drug delivery and physically targeting myc directly given that it is a largely disordered protein that lacks discernible clefts and pockets for small molecules to inhabit. Furthermore, c-myc is important in normal tissue maintenance so the effect of its inhibition in humans is difficult to predict. However, recent in vivo studies showed that systemic myc inhibition (using the peptide pan myc inhibitor Omomyc) has mild and reversible side effects and induces tumour regression. This has alleviated concerns about the side effects that myc inhibition might have, and reinforced the promise of myc as a powerful drug target. However, the translation of Omomyc into the clinic has been hindered by poor cellular delivery. In fact, no direct myc inhibitor has yet been approved, indicating that novel approaches are needed. Moreover, inhibitors in development tend to inhibit all myc family proteins. An inhibitor that could specifically target N-myc might improve safety through bypassing c-myc inhibition. This could be used for the treatment of N-myc-driven cancers such as MYCN-amplified neuroblastoma. Nanobodies, camelid-derived single-domain antibodies, are a relatively new drug class. Whilst some are already in clinical trials for a wide range of diseases, these are specific for cell-surface or extracellular targets. However, their properties also make them ideal for use as intracellular antibodies or ‘intrabodies’. For example, they are small (just 12-15 kDa) and highly soluble due to naturally occurring hydrophobic to hydrophilic amino acid substitutions. Their small size and convex shape makes them advantageous in capturing structures in intrinsically disordered proteins and allows them to reach hidden epitopes not accessible to conventional antibodies, which could improve biological activity. Importantly, nanobodies retain the high specificities and affinities of conventional antibodies. Their small, single-domain nature also means they can be engineered with ease to modify aspects of their localisation and/or function. For example, they can be coupled to carrier molecules to facilitate cellular entry, and a nuclear localisation signal (NLS) can be added to drive them into the nucleus. Also, it was recently shown that an F-box domain could also be incorporated into nanobodies to recruit degradation machinery to its antigen, which depletes the antigen from cells via the proteasomal degradation pathway. Due to their highly advantageous properties, nanobodies raised against N-myc might overcome the barriers to targeting N-myc, providing potent and specific means of directly inhibiting N-myc therapeutically, which has not yet been achieved. In this thesis, nine unique nanobodies were raised against N-myc. These included three against the basic helix-loop-helix leucine zipper (bHLH-LZ) domain where Max dimerises, and six against the transactivation domain where numerous regulatory and cofactor proteins bind, such as the E3 ubiquitin ligase Skp2. Nanobodies against the transactivation domain were more specific for N-myc and were shown to inhibit its Skp-2-mediated ubiquitylation. This could provide novel means of eradicating tumours based on a study showing that inhibition of ubiquitylation at this domain triggers a transcriptional ‘switch’ that induces a non-canonical target gene Egr1, leading to p53-independent apoptosis. A nanobody against the bHLH-LZ (Nb C2) was shown to bind both N- and c-myc to similar magnitudes. Its affinity for N-myc bHLH-LZ was superior to that of the small molecule myc inhibitor 10058-F4, which prolongs survival in a MYCN-dependent mouse model of high-risk neuroblastoma. Nb C2 spontaneously transduced cell membranes and its coupling to a novel small molecule carrier (SMoC) enhanced its cellular uptake. Furthermore, the addition of a NLS increased its nuclear localisation. Preliminary experiments showed that Nb C2 might slow proliferation and induce apoptosis in cancer cell lines expressing c-myc, suggesting that Nb C2 might also be effective against cancers characterised by deregulated c-myc. Taken together, data generated in this thesis have revealed intriguing findings that provide a basis for the development of these nanobodies for the treatment of N-myc- and c-myc-driven cancers.

No description available.

During the past few years a development of breeding llamas and alpacas took place in Czech Republic. Increasing interest in those animals entails the need to determine how they respond to the conditions of our country. The aim of this thesis is to evaluate the growth ability of crias, some of the physical dimensions (withers height) and bodyweight of adult llamas. The same indicators are to be evaluated for alpacas as well. The results are to be compared with the standard for llamas and alpacas. A total of 104 animals were measured and weighted, namely 44 llamas and 60 alpacas, and 280 pairs of weight and height data were obtained. Weighing and measuring of adult llamas and alpacas took place once a year around the end of August for two years (2009 and 2010). Young llamas were weighed and measured at approximately monthly intervals according to the possibilities of their breeders. Young alpacas were weighed at approximately weekly intervals. The thesis is divided into two parts. The first part evaluates llamas. It was discovered that the average birth weight of llamas in the Czech Republic is 11 kg. Once mature, they reach an average live weight of 130.3 kg and average height of 110.2 cm. Average daily gains were observed as follows: from birth to 7 weeks 0.20 kg per day, from 7 weeks to 24 weeks 0.18 kg per day and from 24 to 72 weeks 0.13 kg per day. The second part of this thesis is devoted to alpacas, for which the same parameters as for the llamas were monitored. It was found that the average birth weight of alpacas in the Czech Republic is 6.5 kg. Once mature, they reach an average live weight of 65.6 kg and average height at 88.4 cm. Average daily gains are at the following levels: from birth to 7 weeks 0.1 kg per day, from 7 weeks to 24 weeks 0.09 kg per day and from 24 weeks to 72 weeks 0.08 kg per day. It was found that these two species reach final physical maturity later in the Czech Republic than stated by literature. Compared to the standard, most of the llamas and alpacas are up to their standards.

Seminario de Título entregado a la Universidad de Chile en cumplimiento parcial de los requisitos para optar al Título de Química Ambiental. / Los suelos se han constituido como el principal sumidero de metales pesados y otros contaminantes producto de causas naturales y actividades antropogénicas. Ejemplo de estas actividades son las mineras que liberan metales pesados, tales como Hg+2, Pb+2, Cu+2 y Zn+2, y las plantas petroquímicas que producen hidrocarburos aromáticos como Benceno, Tolueno, Etilbenceno, y Xilenos (BTEX) a partir de la fracción volátil del petróleo. El objetivo de este seminario de título consiste en validar un método analítico para la cuantificación de benceno en microcosmos conformados por muestras de suelo co-contaminado contenidas en viales, a los cuales se le adiciona una bacteria especializada. De esta forma, se desarrolla una metodología para determinar benceno remanente en suelo, carente de solventes orgánicos, simple y costo-efectiva, mediante un sistema de extracción de espacio de cabeza acoplado a un cromatógrafo de gases junto a un detector de ionización de llama (HS-GC-FID), que permita monitorear la cinética de remoción de benceno en un suelo co-contaminado con Hg (II), durante un proceso de biorremediación bacteriana utilizando la cepa modificada genéticamente Cupriavidus metallidurans MSR33 que es altamente resistente a mercurio. De esta manera, en el presente trabajo se utilizó un suelo contaminado con benceno y mercurio como una aproximación a una situación real de co-contaminación, donde se aplicó una técnica de remediación a través de la utilización de bacterias especializadas capaces de remover benceno en presencia de mercurio. La medición de benceno con la metodología validada permitió monitorear la cinética de biorremediación utilizando la bacteria C. metallidurans MSR33 entregando información rápida y veraz al aplicar directamente en viales con microcosmos de suelo contaminados con Benceno 200 mg×Kg-1 y Hg (II) 2 mg×Kg-1. La determinación de benceno permitió demostrar que este proceso de biorremediación conforma una novedosa tecnología costo-efectiva y amigable con el medioambiente, aplicable a suelos impactados con BTEX en presencia de metales tóxicos. / Soils have been established as the main sink for heavy metals and other pollutants due to natural causes and anthropogenic activities. Examples of these are mining activities which release heavy metals, such as Hg+2, Pb+2, Cu+2 and Zn+2, and petrochemical plants which produce aromatic hydrocarbons such as Benzene, Toluene, Ethylbenzene, and Xylene (BTEX) from volatile fractions of petroleum. The aim of this seminar consists of validate an analytical method for the quantification of benzene in microcosms consisting on vials containing co-contaminated soil samples, which are inoculated with a specially adapted bacterial culture. The methodology was developed in order to measure benzene concentrations remaining in soil in an organic solvent-free, simple and cost-effective manner. This was carried by means of a headspace extraction system coupled to a gas chromatograph with flame ionization detector (HS-GC-FID), that allowed to monitor the kinetics of benzene removal in a soil co-contaminated with Hg (II), during the process of bacterial bioremediation with the genetically modified strain Cupriavidus metallidurans MSR33 that is highly resistant to mercury. Thus, soil samples polluted with benzene and mercury were used on this research as a practical approach to a real co-contamination scenario. A remediation technique was applied by means of bacteria specially adapted to remove benzene from soil in presence of mercury. The measurement of benzene with the validated methodology allowed to monitor the kinetics of bioremediation using the C. metallidurans MSR33 bacterial strain delivering fast and accurate information when applied directly in vials with soil microcosms contaminated with Benzene 200 mg×Kg-1 and Hg (II) 2 mg×Kg-1. The determination of benzene allowed to demonstrate that this process of bioremediation forms a novel, cost-effective and environmental-friendly technology, potentially useful for treatment of soils impacted with BTEX in the presence of toxic metals.

Considering the activity of introducing genetic resources with a high potential for yield, and with a wide adaptation range, it is possible to strengthen the cattle production system, through the production of adequate forage in quality, and quantity. Brazilian grass was introduced to Bolivia in 1974 for demonstrating excellent forage characteristics, and great adaptation qualities to the different ecosystems of the Andes. The material was introduced through the Experimental Station of Patacamaya, with their material supplied through the Experimental Station of Obonuco (Colombia). The following study was conducted at the Agropecuary Experimental Center of Condori, located 49 km. north of the city of Oruro, department of Oruro, and 12 km. northeast of the population of Caracollo. Geographically it is located between the parallels 17°31’41’’ latitude south, and 64°14’ 02’’ longitude west to the Greenwich, and at a height of 3830 m.a.s.l during the year of 2005. The study was performed with the objective of determining the apparent digestibility in Lamas feed with Brazilian grass, and with Wheat, for which 4 species between 2 to 3 years and an average weight of 79 were selected for the study. The specimens were selected according to phenotypic characteristics, each of them having their own metabolic cage with its own feeding and drinking station. The design was completely random with no structured treatments; the significant variables were evaluated under contrast test to 0.05 of probability. The obtained results regarding the quantity of food consumed is much related to the quantity of feces excreted without having statistical differences between these two variables, the average consumption being of 2157.8 g/day and the average production of feces of a 789.55 g/day. The consumption of water is statistically different but proportional to the amount of moisture found in the forage, meaning that the greater amount of water supplied through the plan the less the amount of drinking water. The species consuming Brazilian grass presented a water consumption of 244.64 cc/day in relationship to the moisture percentage of 11.51% of the plant, being 2.38% larger in comparison to wheat. The amount of feces produced by the experimental units shows no statistical difference at a 0.05 of probability in relationship to food consumption, meaning that the amount of feces produced is proportional to amount of ingested food. The urine volume registered 1540.32 cc/day in relationship with the consumption of Brazilian grass, and a lesser volume of 671 cc/day in relationship with wheat consumption, observing significance with 99% of reliability. The results of the digestibility of the nutrients in Brazilian grass, and wheat are the following: Protein 78.59% and 48.01%; Ash 72.99% and 52.59%; Fiber 47.70% and 52.51%; Ethereal Extract 62.64% and 53.11%, respectively. We can conclude that the Brazilian grass is an excellent forage specie of good digestible properties, being protein its principal digestible nutrient.

The present work consisted in the use of biochemical and molecular nutrition blocks (BMN) in llamas during the last third of the gestation period, at the Agricultural Experiment Center of Condori (CEAC), located 49 km north of the city of Oruro, and 12 km north of the Caracollo population, in the province of Cercado. For this research work, we used 20 pregnant llamas in the last third of their gestation period, between the ages of 2 and 3 years old, 10 of which belonged to the Khara breed, and 10 to the Thampulli breed. Two corrals of 7 x 7 m., with adobe walls of a height of 1.80 m., a soil floor, and a metallic door, were built for the placement of the animals. The following ingredients were used for the preparation of the BMN blocks: 100 L of molasses, 75 kg of barley flour, 18 kg of alfalfa flour, 12 kg of sorghum flour, 30kg of mineral salt, 30 kg of Urea to the 46%, and 30 kg, of clay. The following variables were considered: Final body weight of the mother (before birth), increment of live weight of the mother by date, body weight of the mother (after birth), body weight of the llama calf, body weight gain of the mother, mother's food consumption, and physical-chemical analysis of the feeding blocks. The variables of the study were analyzed under a completely randomized design test, and for the discrimination of variable averages we used the Duncan test to a 95% of reliability. We came to the following conclusions: The greater increment of live weight was reported by llamas that received the BMV block as a dietary supplement, the gain was of 107.91 Kg/Llama, and the least was reported by those who did not receive the BMV blocks as a dietary supplement with and increment of 102.07 Kg/Llama. For the final live weight of the mothers, the greater number was reported by llamas that received the BMV blocks, reporting 112.0 Kg/Llama, the least was reported by those who did not receive the supplement with an increase of 105.10 Kg/Llama. Post-partum Llamas that received supplementation reported a live weight increase of 97.43 Kg/Llama, whereas those who did not receive the supplement reported an increase of 88.40 Kg/Llama. According to breed the Khara Llama reported an increase of 96.56 Kg/llama, while the Thampulli line reported an increase of 89.10 Kg/Llama. The greater live weight of the calf at birth was reported by those whose mothers were given the BMN supplements; reporting 11.50 Kg/Calf, while those whose mothers did not receive the supplement reported 9.60 Kg/ Calf. Therefore, we recommend the supplementation of the diet with biochemical and molecular nutrition blocks in the feeding of llamas in the last third of the gestation period to obtain calves with higher body weight at birth.

At an international level, Bolivia possesses the largest population of llamas followed by Peru. This makes our country a power, in this renewable resource, to create and generate economic development primarily for the quality of llama meat which has nutritional properties superior to other species of animals used for food. The development of the investigation of llamas in our country has not been very far-reaching. In other countries, advances have been made for quite some time, specifically in the area of reproduction, which presents problems in bringing about an intensive repopulation due to the unique characteristics of llama copulation and semen. With the present work, carried out in the Unidad Académica Campesina - Tiahuanaco (Universidad Católica Boliviana), titled: Determination of the Protein Fraction and Presence of Antibodies in the Seminal Fluid of Llamas of 3, 4, and 5 years of age, in Different Dates of Semen Collection, the number of protein fractions, their molecular weights, the presence of antibodies, and the concentrations of the protein fractions was determined. This will lead to determining the possible presence of a component in the seminal fluid which is responsible for the low percentage of births achieved in these animals. In studies conducted in the field, a series of tests were carried out so the male would respond to the artificial collection of the semen, beginning with the puppet technique (Peruvian) and others, all of which did not produce results. The objective was achieved with the mannequin of a llama's hindquarters which was accepted by 80% of the males. Laboratory studies consisted in separating seminal fluid from the semen through centrifugation. An electrophoresis technique was employed in order to examine the protein fractions. This technique allowed determination of the number of protein fractions, their molecular weights, immunoglobulin, and their concentrations. The highest number of protein fractions was found in 4 and 5-year-old animals with 15 to 18 protein fractions. In the weekly collection of semen, the final week returned 18 protein fractions (in the 4-year-old animal), which fractions may possibly play an important role in the nutrition, capacitation, and protection of the spermatozoid. The molecular weights of the proteins found in the seminal fluid vary from one animal to another and in the collection dates. The extremes in the range of molecular weights are from 2,000 to 150,000 Daltons. Animals five years in age presented higher molecular weights compared with younger animals. Regarding immunoglobulin or antibodies, proteins were found with molecular weights from 150,000 to 152,500 Daltons in animals from three to five years of age. Through the radial immunodiffusion technique, immunoglobulin G was found in the llama seminal fluid. The protein fraction concentrations are variable, ranging from 11.65 to 0.03 mg/mL of seminal fluid. These variations are heterogeneous as much in animal age as in dates of collection. There exists a loose superiority in five-year-olds and during the third week of semen collection. The obtained results of the protein fractions and their molecular weights demonstrate the existence of antibodies in the seminal fluid of the llama that play an important role in spermatic viability. Because of this, continued investigation should be carried out to determine the antibodies' specific function and find alternatives to isolate and neutralize them and increase the fertility figures of these animals.

Sensitive data leaking from a system can have tremendous negative consequences, such as discrimination, social stigma, and fraudulent economic consequences for those whose data has been leaked. Therefore, it’s of utmost importance that sensitive data is not leaked from a system. This thesis investigated different methods to prevent sensitive patient data from leaking in a machine learning system. Various methods have been investigated and evaluated based on previous research; the methods used in this thesis are a large language model (LLM) for code analysis and a membership inference attack on models to test their privacy level. The LLM code analysis results show that the Llama 3 (an LLM) model had an accuracy of 90% in identifying malicious code that attempts to steal sensitive patient data. The model analysis can evaluate and determine membership inference of sensitive patient data used for training in machine learning models, which is essential for determining data leakage a machine learning model can pose in machine learning systems. Further studies in increasing the deterministic and formatting of the LLM‘s responses must be investigated to ensure the robustness of the security system that utilizes LLMs before it can be deployed in a production environment. Further studies of the model analysis can apply a wider variety of evaluations, such as increased size of machine learning model types and increased range of attack testing types of machine learning models, which can be implemented into machine learning systems. / Känsliga data som läcker från ett system kan ha enorma negativa konsekvenser, såsom diskriminering, social stigmatisering och negativa ekonomiska konsekvenser för dem vars data har läckt ut. Därför är det av yttersta vikt att känsliga data inte läcker från ett system. Denna avhandling undersökte olika metoder för att förhindra att känsliga patientdata läcker ut ur ett maskininlärningssystem. Olika metoder har undersökts och utvärderats baserat på tidigare forskning; metoderna som användes i denna avhandling är en stor språkmodell (LLM) för kodanalys och en medlemskapsinfiltrationsattack på maskininlärnings (ML) modeller för att testa modellernas integritetsnivå. Kodanalysresultaten från LLM visar att modellen Llama 3 hade en noggrannhet på 90% i att identifiera skadlig kod som försöker stjäla känsliga patientdata. Modellanalysen kan utvärdera och bestämma medlemskap av känsliga patientdata som används för träning i maskininlärningsmodeller, vilket är avgörande för att bestämma den dataläckage som en maskininlärningsmodell kan exponera. Ytterligare studier för att öka determinismen och formateringen av LLM:s svar måste undersökas för att säkerställa robustheten i säkerhetssystemet som använder LLM:er innan det kan driftsättas i en produktionsmiljö. Vidare studier av modellanalysen kan tillämpa ytterligare bredd av utvärderingar, såsom ökad storlek på maskininlärningsmodelltyper och ökat utbud av attacktesttyper av maskininlärningsmodeller som kan implementeras i maskininlärningssystem.

Vikten av hållbarhetsredovisning kan ses genom den uppmärksamhet ämnet har från företag, media, myndigheter och den ökande regleringen genom införandet av nya direktiv och lagstiftning. Att manuellt analysera företags hållbarhetsredovisningar är en tidskrävande process. En automatiserad analys av hållbarhetsredovisningar skulle innebära ekonomiska och tidsmässiga vinster när viktiga insikter tas fram relaterat till större företags påverkan på sin miljö och omgivning. Denna studie syftar till att utforska möjligheterna till en automatisering av en befintlig manuell arbetsmetod. Prototypen som utvecklats tillämpar moderna språkbehandlingsmetoder, ett område inom maskininlärning, för att realisera denna vision. Studiens implementation uppnår för de utvärderade språkmodellerna upp till 96% precision för majoritetsklassen vid bearbetning av grunddatat respektive 55% precision för minoritetsdataklassen vid bearbetning av grunddata jämfört resultat från den manuellt genomförda metoden. Slutsatsen är att en automatiserad version av den befintliga manuella analysmetoden kan konstrueras och även förbättras med den snabba utveckling som sker inom teknologi och språkmodeller, om ytterligare resurser avsätts. Resultaten visar hopp om potentialen för en metodik som utvecklas i vidare arbeten. / The importance of sustainability reporting can be observed by the attention directed towards the subject from companies, media and authorities’ continuous new directives and laws. To manually analyze companies’ sustainability reports is a time-consuming process. An automated approach analyzing sustainability reports would give advantages regarding both time and economics when important insights related to companies’ operations are brought into light. This study aims to explore possibilities in automating an existing manual method related to analyzing sustainability reports. The developed prototype applies modern language models and methods related to machine learning to realize this vision. For the evaluated language models, the study’s implementation achieves up to 96% precision for the majority class, while the minority class achieves up to 55% precision in processing of data, when compared to reference results from the manual evaluation method. The work’s conclusion indicates that an automated version of the existing manual method for analysis can be constructed with sufficient resources, and even further improved as the area of technology further advances. The results are positive for the potential for a more sophisticated method that can be developed in further work.