[EN] Domain-specific languages (DSLs) are considered to be a powerful tool for enhancing the efficiency of software developers and bring software development closer to end-users from complex domains. However, the successful development of a DSL for a complex domain is a challenge from the technical point of view and because end-user acceptance is key. Despite this fact, the relevant role of end-users during DSL development has traditionally been neglected. Normally, end-users participate at the beginning to communicate their preferences but they do not participate again until the DSL is completely implemented. As a consequence, if the language to develop reaches a complex domain, the chances that errors appear in the DSL are higher and solving them could involve large modifications that could have been avoided. As a solution, in this PhD thesis, we propose an agile, model-driven method to involve end-users in DSL development. This thesis researches if the combination of best practices from the model-driven development (MDD) discipline and best practices from agile methods is a suitable approach to involve end-users in the DSL development process. In order to validate the proposal, we have selected a highly complex domain such as the genetic analysis domain and we have collaborated with geneticists from three organizations. The proposed method has been used to involve these geneticists in the development of a DSL for the creation of genetic analysis pipelines. Simultaneously, we have carried out an empirical experiment to validate whether end-users and developers were satisfied with the proposal. / [ES] Los lenguajes específicos de dominio (DSLs) son una herramienta muy potente para mejorar la eficiencia de los desarrolladores de software, así como para acercar el desarrollo software a usuarios sin conocimientos informáticos. Sin embargo, su principal problema es que desarrollar un DSL es complejo; no sólo desde el punto de vista técnico, sino especialmente porque la aceptación de dicho lenguaje por parte de los usuarios finales es clave. A pesar de este hecho, los métodos tradicionales de desarrollo de DSLs no enfatizan el importante rol de los usuarios finales durante el desarrollo. Normalmente, los usuarios participan al inicio para comunicar sus preferencias, pero no vuelven a participar hasta que el DSL está completamente desarrollado. Si el lenguaje a desarrollar aborda un dominio complejo, la posibilidad de que existan errores en el DSL es mayor, y su solución podría conllevar a modificaciones de gran calibre que podrían haberse evitado. Como solución, en esta tesis proponemos un método de desarrollo de DSLs, ágil, y dirigido por modelos que involucra a los usuarios finales. Esta tesis investiga si la combinación de buenas prácticas del desarrollo dirigido por modelos (MDD) y de buenas prácticas de métodos ágiles es adecuada para involucrar a los usuarios finales en el desarrollo de DSLs. Para validar la idoneidad de la propuesta, se ha seleccionado un dominio complejo como el de los análisis genéticos y se ha colaborado con un conjunto de genetistas procedentes de tres organizaciones. El método propuesto se ha utilizado para involucrar a dichos genetistas en el desarrollo de un DSL para la creación de pipelines para el análisis genético. Conjuntamente, se ha llevado a cabo un experimento empírico para validar si los usuarios finales y los desarrolladores están satisfechos con la propuesta de la presente tesis. En resumen, las contribuciones principales de esta tesis doctoral son el diseño e implementación de un método innovador, ágil y dirigido por modelos para involucrar a los usuarios finales en el desarrollo de DSLs, así como la validación de dicha propuesta en un entorno industrial en un desarrollo real de un DSL. / [CA] Els llenguatges específics de domini (DSLs) son una ferramenta molt potent per a millorar l'eficiència dels desenvolupadors de programari, així com per a apropar el desenvolupament de programari a usuaris sense coneixements informàtics. El problema es que desenvolupar un DSL es complex, no sols des del punt de vista tècnic, sinó especialment perquè l'acceptació de dit llenguatge per part dels usuaris finals es clau. Malgrat aquest fet, els mètodes tradicionals de desenvolupament de DSLs no emfatitzen l'important rol dels usuaris finals durant el desenvolupament. Normalment, els usuaris participen a l'inici per a comunicar les seues preferències, però no tornen a participar fins que el DSL està completament desenvolupat. Si el llenguatge a desenvolupar aborda un domini complex, la possibilitat de que hi hagen errors en el DSL es major i solucionar-los podria implicar modificacions de gran calibre que podrien haver-se evitat. Com a solució, en aquesta tesis proposem un mètode de desenvolupament de DSLs, àgil i dirigit per models que involucra als usuaris finals. Aquesta tesis investiga si la combinació de bones pràctiques del desenvolupament dirigit per models (MDD) i de bones pràctiques de mètodes àgils es adequada per a involucrar els usuaris finals en el desenvolupament de DSLs. Per a validar la idoneïtat de la proposta, s'ha seleccionat un domini complex com el dels anàlisis genètics i s'ha col·laborat amb un conjunt de genetistes procedents de tres organitzacions. El mètode s'ha utilitzat per a involucrar a dits genetistes en el desenvolupament d'un DSL per a la creació de pipelines per al anàlisis genètic. Al mateix temps, s'ha dut a terme un experiment empíric per a validar si tant els usuaris finals com els desenvolupadors estan satisfets amb la proposta de la present tesis. En resum, les contribucions principals d'aquesta tesis doctoral son el disseny i implementació d'un mètode innovador, àgil i dirigit per models per a involucrar als usuaris finals en el desenvolupament de DSLs, així com la validació de la proposta en un entorn industrial amb un desenvolupament real d'un DSL. / Villanueva Del Pozo, MJ. (2016). An agile model-driven method for involving end-users in DSL development [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/60156

Xylella fastidiosa (Xf) e Xanthomonas axonopodis pv. citri (Xac) são gama proteobactérias gram negativas, responsáveis por grandes perdas econômicas no setor citrícola brasileiro. Com seus genomas seqüenciados e anotados, fizemos uma análise comparativa entre suas composições gênicas e seus ambientes de vida. Xac apresenta um genoma de 5.2Mb contra 2.7Mb de Xf Isto reflete no número de genes (4432 contra 2838) que acabam refletindo em uma maior complexidade metabólica de Xac, caracterizada por: uma extensa gama de genes de degradação de parede celular (44), biossíntese de proteases (92), genes de funções regulatórias (296), um completo metabolismo energético (209), quimiotático (inexistente em Xf) e secretório (presença dos tipos I, II, III e IV , sendo o II em duplicata), além de um grande número de genes envolvidos com captação de ferro (65), fazem de Xac um patógeno de alto poder invasivo e de rápida propagação e virulência Em contrapartida, Xf por não possuir a complexidade supracitada, parece ter seus recursos adaptados ao ambiente em que vive, como por exemplo um alto número de genes envolvidos com biossíntese de pili, que associado à biossíntese de goma, favorecem sua adesão nas glândulas salivares do vetor (cigarrinha) e a formação de aglomerados celulares responsáveis pelo entupimento dos vasos que levam às patologias decorrentes do evento. / Xylella fastidiosa (Xf) and Xanthomonas axonopodis pv. citri Xac are gram negative gamma proteobacteria, responsible for great economical losses in the Brazilian citrus sector. With their sequenced and annotated genomes, we have done a comparative analysis between their genetic composition and life habitat. Xac displays a genome of 5.2Mb against 2. 7Mb of Xf. This reflects the number of genes (4432 against 2838) which results in a greater metabolic complexity of Xac, characterized by: a wide range of genes of cell wall degradation (44), biosynthesis of proteases (92), many genes of regulatory functions (296), a complex energy metabolism (209), chemotatic (absent in Xf) and secretory systerns (presence of types I, II, III and IV, type II in duplicate), besides a great number of genes involved in iron acquisition (65), make of Xac a pathogen of high invasive power and of quick spreading and virulence. In the other hand Xf, due to the lack of the complexity just cited, seems to have its resources adapted to the habitat in which it lives, as for example a large number of genes involved in pili biosynthesis, that associated with gum biosynthesis, favor its adhesion to the salivary glands of the vector (sharpshooter) and the formation of cellular agglomerations responsible for the blockage of the vessels which leads to the pathologies resulted from this event.

Xylella fastidiosa (Xf) e Xanthomonas axonopodis pv. citri (Xac) são gama proteobactérias gram negativas, responsáveis por grandes perdas econômicas no setor citrícola brasileiro. Com seus genomas seqüenciados e anotados, fizemos uma análise comparativa entre suas composições gênicas e seus ambientes de vida. Xac apresenta um genoma de 5.2Mb contra 2.7Mb de Xf Isto reflete no número de genes (4432 contra 2838) que acabam refletindo em uma maior complexidade metabólica de Xac, caracterizada por: uma extensa gama de genes de degradação de parede celular (44), biossíntese de proteases (92), genes de funções regulatórias (296), um completo metabolismo energético (209), quimiotático (inexistente em Xf) e secretório (presença dos tipos I, II, III e IV , sendo o II em duplicata), além de um grande número de genes envolvidos com captação de ferro (65), fazem de Xac um patógeno de alto poder invasivo e de rápida propagação e virulência Em contrapartida, Xf por não possuir a complexidade supracitada, parece ter seus recursos adaptados ao ambiente em que vive, como por exemplo um alto número de genes envolvidos com biossíntese de pili, que associado à biossíntese de goma, favorecem sua adesão nas glândulas salivares do vetor (cigarrinha) e a formação de aglomerados celulares responsáveis pelo entupimento dos vasos que levam às patologias decorrentes do evento. / Xylella fastidiosa (Xf) and Xanthomonas axonopodis pv. citri Xac are gram negative gamma proteobacteria, responsible for great economical losses in the Brazilian citrus sector. With their sequenced and annotated genomes, we have done a comparative analysis between their genetic composition and life habitat. Xac displays a genome of 5.2Mb against 2. 7Mb of Xf. This reflects the number of genes (4432 against 2838) which results in a greater metabolic complexity of Xac, characterized by: a wide range of genes of cell wall degradation (44), biosynthesis of proteases (92), many genes of regulatory functions (296), a complex energy metabolism (209), chemotatic (absent in Xf) and secretory systerns (presence of types I, II, III and IV, type II in duplicate), besides a great number of genes involved in iron acquisition (65), make of Xac a pathogen of high invasive power and of quick spreading and virulence. In the other hand Xf, due to the lack of the complexity just cited, seems to have its resources adapted to the habitat in which it lives, as for example a large number of genes involved in pili biosynthesis, that associated with gum biosynthesis, favor its adhesion to the salivary glands of the vector (sharpshooter) and the formation of cellular agglomerations responsible for the blockage of the vessels which leads to the pathologies resulted from this event.

<p>Morfometrijske analize su ra&ntilde;ena merenjem krilne nervature 540 uzoraka krila<br />pčela sa 9 lokaliteta u Vojvodini. Izračunato je 16 uglova (A1, A4, B3, B4, D7, E9, G7,<br />G18, H12, J10, J16, K19, L13, M17, O26, Q21) koje zaklapa krilna nervatura i 4 indeksa<br />(Ci, Pci, Dbi, Ri), ukupno 20 mera. Izračunate su prosečne vrednosti i utvr&ntilde;ena je<br />statistička značajnost razlika izme&ntilde;u pčela iz regona Srema i Bačke i pčela iz regiona<br />Banata, tako&ntilde;e je izvr&scaron;eno i pore&ntilde;enje pčela svih lokaliteta sa DAWINO standardima za<br />5 rasa pčela (Apis mellifera carnica, Apis mellifera macedonica, Apis mellifera mellifera,<br />Apis mellifera ligustica i Apis mellifera caucasica).<br />Analizom varijanse izračunatih 20 osobina krilne nervature, utvr&ntilde;eno je da samo<br />kod osobine A4 nisu utvr&ntilde;ene statistički značajnie razlike izme&ntilde;u posmatranih<br />lokaliteta, dok su u 19 osobina utvr&ntilde;ene statistički značajne razlike.<br />Utvr&ntilde;ivanjem statističke značajnosti razlika, pčela iz regiona Srema i Bačke i pčela<br />iz regiona Banata, utvr&ntilde;eno je da 45% osobina ne pokazuju statistički značajne razlike,<br />dok 45% osobina pokazuje statistički vrlo značajne razlike (P&lt;0,01) i 10% osobina<br />pokazuje statistički značajne razlike (P&lt;0,05).<br />Upore&ntilde;ivanjem dobijenih vrednosti 20 parametera krilne nervature, pomoću ztesta,<br />sa DAWINO standardima za pet rasa pčela, utvr&ntilde;eno je da na bazi celog uzorka<br />statistički nema značajnih razlika kod osobina A4 i D7 sa A. m. carnica, kod osobina<br />H12, G18 i B4 sa A. m.macedonica i kod osobina J16 i B4 pore&ntilde;eno sa rasom A.<br />m.ligustica. Kod pčela iz regiona Srema i Bačke utvr&ntilde;eno je da statistički nema značajnih<br />razlika kod osobina A4, B3, D7 i G18 upore&ntilde;eno sa A.m. carnica, kod osobina H12 i B4<br />upore&ntilde;eno sa A. m.macedonica i kod osobina G18, K19, J16 i Q21 upore&ntilde;eno sa rasom<br />A. m.ligustica, dok kod pčela iz regiona Banata utvr&ntilde;eno je da statistički nema značajnih<br />razlika kod osobina A4, E9, D7 i J10 upore&ntilde;eno sa A.m. carnica, kod osobina H12, J10,<br />L13 i PCi upore&ntilde;eno sa A. m.macedonica i kod osobina B4, J16 i PCi upore&ntilde;eno sa<br />rasom A. m.ligustica.<br />Ocena genetičke povezanosti, unutar populacijska raznolikost i struktura<br />populacije, dva tipa pčela u Vojvodini, izračunata je na bazi varijacije alela 25 lokusa<br />mikrosatelita. Izvr&scaron;ena je genetska tipizacija sledećih mikrosatelita: A8, A14, A24, A29,<br />A43, A79, A88, A113, Ac11, Ac88, Ac139, Ac306, Ap15, Ap68, Ap85, Ap90, Ap223,<br />Ap224, Ap226, Ap249, Ap273, Ap274, Ap288, At168, At188. 92% ili 23 lokusa su se<br />pokazali kao polimorfni u uzorcima pčela iz Srema i Bačke, a 88% ili 22 lokusa su se<br />pokazali kao polimorfni u uzorcima pčela iz Banata. Izračunata heterozigotnost na nivou<br />cele populacije se nije statistički značajno razlikovala od očekivane heterozigotnosti.<br />Utvr&ntilde;eno je da dobijene genetičke razlike izme&ntilde;u analiziranih pčela iz regiona Srema i<br />Bačke i retgiona Banata nisu dovoljne da se ove dve populacije mogu smatrati<br />razdvojenim.</p> / <p> Morphometric analyses have been done by measuring the wing nervature in<br /> 540 samples of bees, collected from nine localities in Vojvodina. 16 angles<br /> formed by wing nervation have been calculated(A1, A4, B3, B4, D7, E9, G7,<br /> G18, H12, J10, J16, K19, L13, M17, O26, Q21) as well as four indexes (Ci, PCI,<br /> DBI , R), a total of 20 measures. The average values have been calculated and<br /> statistical significant differences in bees from Srem, Backa and Banat region<br /> determined. Five breeds of bees from these regions have been compared to<br /> Dawino standards.<br /> The analyses of the variance of calculated 20 features of wing nervature indicate<br /> that statistically significant differences in monitored localities have not been found<br /> only in A4, on the other hand in 19 properties significant differences have been<br /> discovered.<br /> Established statistically significant differences between breeds from Srem<br /> and Backa regions reveale that 45% properties do not show any statistically<br /> important differences, while 45% features show very important statistical<br /> differences (P&lt;0,01) and 10% show statistically important differences (P&lt;0,05).<br /> It has been established by comparing the obtained values of 20 parametres<br /> of wing nervature by means of z test to DAWINO standards for five breeds of<br /> bees that, based on the whole sample, there are no significant differences in<br /> features A4 and D7 in A.m. carnica, in features H12, G18 and B4 in A.m.<br /> macedonica and features J16 and B4 compared to A.m. ligustica. As for bees from<br /> Srem and Backa region,there are statistically no significante differences in<br /> features A4, B3, D7 and G18 compared to A.m. carnica, features H12 and B4<br /> compared to A.m. macedonica and features G18, K19, J16 and Q21 compared to<br /> A.m. ligustica, while in bees from Banat region, statistically there are no<br /> significant differences in features A4, E9, D7 and J10 compared to A.m. carnica,<br /> features H12, J10, L13 and Pci compared to A.m. macedonica and features B4,<br /> J16 and Pci compared to A.m. ligustica.<br /> The evaluation of genetic correlation, the diversity of bees population and<br /> population structure of two types of bees in Vojvodina have been established on<br /> the basis of allels variations in 25 locus microsatelites.The following<br /> microsatelites have been standardized &ndash; A8,A14,A24,A29, A43, A79, A88, A113,<br /> Ac11, Ac88, Ac139, Ac306, Ap15, Ap68, Ap85, Ap90, Ap223, Ap224, Ap226,<br /> Ap249, Ap273, Ap274, Ap288, At168, At188. 92% or 23 locus have shown as<br /> polymorphs in bees from Srem and Backa and 88% or 22 locus samples have<br /> shown as polzmorphs in bees samples from Banat and Backa region. The whole<br /> population calculated heterozygosity has not shown statistically significant<br /> differrence from expected heterozygisity. It has been established that the obtained<br /> genetic differences between the analysed bees from Srem and Backa region and<br /> Banat region are not significant to indicate two populations.</p>

<p>Gilbert’s syndrome (GS) occurs in approximately 10% of the European population. The most common cause is homozygosity for UGT1A1*28, which is a TA repeat expansion in the promoter of UGT1A1. It is characterised by intermittent hyperbilirubinemia due to reduced hepatic activity of the  enzyme UDP-glucuronosyl-transferase 1A1(UGT1A1). GS also  alteres the pharmacokinetics of some drugs and increases the risk of drug toxicity. Irinotecan (Camptosar®, Campto®) is used in metastatic colorectal cancer and the active metabolite is inactivated by UGT1A1. Studies have shown that GS can be a risk factor for toxicity during irinotecan therapy.</p><p>Three different methods for genotyping of UGT1A1*28 have been tested.</p><p>PCR with electrophoresis used for size separation, melting temperature analysis and fluorescent PCR followed by fragment analysis on a capillary sequencer.</p><p>The last method was found to be superior. This method was used for genotyping of patients with colorectal cancer treated with irinotecan and 5-fluorouracil in the Nordic VI study. A significant association between UGT1A1 genotype and plasma bilirubin level before the start of irinotecan treatment was seen (ANOVA p<0.0001). Patients with GS had an overall increased risk of adverse drug reactions (Fishers Exact test p=0.02).</p><p>Gilbert’s syndrome can be diagnosed by genotyping UGT1A1*28 with a fragment analysis method. Genotyping of UGT1A1*28 can be used to identify patients with an increased risk of adverse reactions to irinotecan.<strong> </strong></p><p><strong> </strong></p> / <p>Gilberts syndrom (GS) drabbar upp till 10% av befolkningen i Västeuropa. GS beror på nedsatt aktivitet av enzymet UDP-glukuronosyltransferas 1A1 (UGT1A1) i levern. Den vanligaste orsaken är att individen är homozygot för en insertion av två baser i promotorn för genen UGT1A1. Denna genvariant kallas (TA)7TAA  eller UGT1A1*28. GS leder till intermittent stegring av bilirubin vid infektioner, men bilirubinstegring kan förekomma även utan utlösande agens. GS kan också leda till bilirubinstegring vid viss läkemedelsbehandling. Irinotekan (Campto®) används vid metastaserande colorektal cancer och dess aktiva metabolit inaktiveras av UGT1A1. Det finns rapporter om att GS ger ökad risk för toxiska biverkningar av irinotekan.</p><p>Tre metoder för att bestämma UGT1A1 har jämförts: PCR med elfores, PCR med smältpunktsanalys och PCR med fragmentanalys på sekvensator. Den sista metoden var bäst och användes för att genotypa UGT1A1 hos patienter med colorektal cancer från Nordic VI-studien. De behandlades med irinotekan i kombination med bolusinjektion eller infusion av 5-fluorouracil. Vi fann att  patienter med GS hade signifikant högre S-bilirubin före behandling jämfört med övriga patienter. De hade även ökad frekvens biverkningar av irinotekan (Fishers exakta test p=0,02).</p><p>Genotypning av UGT1A1 kan således användas för att diagnostisera Gilberts syndrom hos patienter med oförklarad bilirubinstegring. Det kan även användas för att identifiera patienter med ökad risk för biverkningar av irinotekan.</p>

Malaria, an infectious disease caused by Plasmodium parasites, is one of the major world-scale health problems. Despite the efforts aimed at finding an effective way to control the disease, the success has been thwarted by the emergence of parasite drug resistance and mosquito resistance to insecticides. This thesis focuses on the genetic analysis of resistance to murine malaria induced by the lethal Plasmodium berghei ANKA using a wild-derived-inbred strain (WDIS). The aim of this thesis was to exploit the genetic diversity represented among WDIS for identifying loci contributing to resistance/susceptibility to murine malaria. The work included a genome-wide polymorphism survey using microsatellite markers performed on 10 WDIS. Comparisons of these strains to laboratory inbred strains confirmed a higher rate of polymorphism among the WDIS. We conclude that these WDIS represent repositories of unique naturally occurring genetic variability that may prove to be invaluable for the study of complex phenotypes. Next, we used the WDIS to search for novel phenotypes related to malaria pathogenesis. Whereas most laboratory strains were susceptible to experimental cerebral malaria (ECM) after infection with P. berghei ANKA, several WDIS were found to be resistant. To study the genetic inheritance of resistant/susceptibility to P. berghei ANKA infection we analysed backcross and F2 cohorts derived from crossing the WLA wild-derived strain with a laboratory mouse strain (C57BL/6). A novel phenotype represented by the cure of infection, clearance of parasitaemia and establishment of immunological memory was observed in the F2 progeny. The backcross progeny was used to genetically map one locus on chromosome 1 (Berr1) and one locus on chromosome 11 (Berr2) that mediate control of resistance to ECM induced by P. berghei ANKA. Genetic mapping using the F2 progeny showed that a locus on chromosome 1 (Berr1) and a locus on chromosome 9 (Berr3) were contributing to control survival time after infection with lethal Plasmodium. Finally, we identified, a locus on chromosome 4 (Berr4) that appears to control time of death due to hyperparasitaemia. This thesis underlines the value of using WDIS to reveal genetic factors involved in the aetiology of disease phenotypes. The characterisation of the genetic factors represented by the malaria resistance loci identified here are expected to provide a better understanding of the malaria pathology.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Iron is an essential element that plays a role in the process of respiration, oxygen transport and as a principle cofactor to several enzymes. Iron homeostasis is a finely regulated process since excess levels become toxic to healthy cells via the production of reactive oxygen species. A plethora of genes that control several key points throughout this regulatory process have been identified. Research focusing on changes in expression levels and downstream functional effects of these genes has become increasingly important over the past decade. One area of particular interest has emerged since a link between iron status and host response to Mycobacterium tuberculosis infection was discovered. Although the prevalence of Tuberculosis has decreased across the globe with the exception of Africa and parts of Europe, the mortality rate remains high. Therefore, research that focuses on understanding an individual’s predetermined susceptibility to TB infection at the genetic level could provide health care practitioners with the tools required to identify and educate at-risk individuals prior to TB infection. RT-qPCR was utilised to determine expression profiles for eight iron genes (CP, CYBRD1, FTH, FTL, LTF, HFE, HMOX1, and SCL40A1) normalised to three reference genes (ACTB, GUSB, and RPL37A1). Up-regulation is demonstrated in the TB group for transcript levels recorded for CYBRD1, HFE, HMOX1, and SLC40A1. Several measured serum parameters including conjugated, unconjugated, total bilirubin, and total protein were increased in the TB group while albumin was significantly lower in this group. Correlation analysis demonstrated that a positive correlation exists between transferrin saturation and iron and a negative correlation exists between transferrin and ferritin levels. Individuals categorised with low serum iron levels demonstrated lower CP/GUSB levels and higher HMOX1/GUSB levels. Individuals categorised with low transferrin saturation levels demonstrated higher FTL/GUSB and SLC40A1/GUSB levels and lower CP/GUSB. Results from this study provide further evidence for the relationship between iron status and TB infection rates, although protein studies are required to confirm these results. The data obtained illustrate the important role that these profiles and iron parameters may play in the clinical field when identifying at-risk individuals. Further investigation that focuses on which gene profile and parameter combinations show the most distinctive utility in the clinical setting is warranted. / AFRIKAANSE OPSOMMING: Yster is ‘n noodsaaklike element wat ‘n rol speel in die proses van respirasie en die vervoer van suurstof en ook ‘n belangrike ko-faktor vir verskeie ensieme is. Yster homeostase is op ‘n fyn manier gereguleer omdat oormatige vlakke toksies kan wees vir gesonde selle wanneer reaktiewe suurstofspesies geproduseer word. ‘n Magdom gene wat verskeie sleutelpunte in hierdie proses kontroleer is voorheen identifiseer. Navorsing wat fokus op die veranderinge in geenuitdrukkingsvlakke en die funksionele gevolge daarvan het oor die afgelope dekade toenemend belangrik geword. Een gebied van spesifieke belang het na vore gekom nadat ‘n verband tussen ystervlakke en die manier waarop die immuunstelsel reageer op Mycobacterium tuberculosis infeksie, ontdek is. Alhoewel die voorkoms van Tuberkulose wêreldwyd, behalwe in Afrika en sekere dele van Europa, afgeneem het, bly die sterftesyfer hoog. Daarom kan navorsing wat daarop fokus om ‘n individu se voorafbepaalde vatbaarheid vir TB-infeksie op die genetiese vlak te verstaan dalk aan gesondheidswerkers die regte instrumente verskaf om hoë-risiko individue te identifiseer en op te voed voordat hulle TB ontwikkel. RT-qPKR is gebruik om die geenuitdrukkingsvlakke van agt ystergene, wat met drie verwysings-gene (ACTB, GUSB, en RPL37A1) genormaliseer is, te bepaal. ‘n Toename in die uitdrukkingsvlakke van CYBRD1, HFE, HMOX1, en SLC40A1 is in die TB-groep waargeneem. Die bloedvlakke van verskeie parameters insluitend gekonjugeerde, ongekonjugeerde, totale bilirubin, en totale proteïen was hoër in die TB-groep, terwyl albuminvlakke laer was in hierdie groep. Korrelasie-analise het ‘n positiewe korrelasie tussen transferrin-versadiging en yster getoon, terwyl daar ‘n negatiewe korrelasie tussen transferrin- en ferritinvlakke gevind is. Individue met lae ystervlakke het laer CP/GUSB-vlakke en hoër HMOX1/GUSB-vlakke getoon. Individue met lae transferrin-versadiging het hoër FTL/GUSB- en SLC40A1/GUSB-vlakke en laer CP/GUSB-vlakke getoon. Resultate uit hierdie studie verskaf verdere getuienis dat daar ‘n verwantskap tussen ystervlakke en TB-infeksiekoerse bestaan, alhoewel proteïenstudies nodig is om hierdie resultate te bevestig. Die data dui op die belangrike rol wat hierdie profiele en ystervlakke in die kliniese veld mag speel in die identifisering van hoë-risiko individue. Verdere ondersoek, gefokus op watter geenprofiel en parameterkombinasies die grootste nut in die kliniese omgewing bied, is geregverdig.

New ideas and methodologies need to be developed to advance our knowledge in the understanding of moral development. The intertwined nature of human activities, communication processes, and the numerous aspects of morality pose a challenge to researchers to construct a methodology that takes into account cognition, affect, sociocultural processes and characteristics, as well as the active role of individuals in their own development. In this paper we aim at suggesting fresh theoretical ideas and a micro genetic methodology to study moral development, particularly within educational contexts. / Se parte de la importancia del desarrollo de nueva ideas y metodologías que posibiliten elavance en la interpretación científica del desarrollo moral. La relación entre la ctividad humana, los procesos de comunicación y las cuestiones de moralidad, colocan a los investigadores frente al desafío de desarrollar un abordaje metodológico del desarrollo moral a partir de un enfoque que considere los aspectos cognitivos, afectivos y socioculturales, así como el papel activo de la persona en su proceso de desarrollo. A partir de esas ideas, nuestro objetivo es proponer el análisis microgenético para el estudio del desarrollo moral, tanto desde el punto de vista teórico como metodológico, en el contexto educacional.

Gilbert’s syndrome (GS) occurs in approximately 10% of the European population. The most common cause is homozygosity for UGT1A1*28, which is a TA repeat expansion in the promoter of UGT1A1. It is characterised by intermittent hyperbilirubinemia due to reduced hepatic activity of the  enzyme UDP-glucuronosyl-transferase 1A1(UGT1A1). GS also  alteres the pharmacokinetics of some drugs and increases the risk of drug toxicity. Irinotecan (Camptosar®, Campto®) is used in metastatic colorectal cancer and the active metabolite is inactivated by UGT1A1. Studies have shown that GS can be a risk factor for toxicity during irinotecan therapy. Three different methods for genotyping of UGT1A1*28 have been tested. PCR with electrophoresis used for size separation, melting temperature analysis and fluorescent PCR followed by fragment analysis on a capillary sequencer. The last method was found to be superior. This method was used for genotyping of patients with colorectal cancer treated with irinotecan and 5-fluorouracil in the Nordic VI study. A significant association between UGT1A1 genotype and plasma bilirubin level before the start of irinotecan treatment was seen (ANOVA p&lt;0.0001). Patients with GS had an overall increased risk of adverse drug reactions (Fishers Exact test p=0.02). Gilbert’s syndrome can be diagnosed by genotyping UGT1A1*28 with a fragment analysis method. Genotyping of UGT1A1*28 can be used to identify patients with an increased risk of adverse reactions to irinotecan. / Gilberts syndrom (GS) drabbar upp till 10% av befolkningen i Västeuropa. GS beror på nedsatt aktivitet av enzymet UDP-glukuronosyltransferas 1A1 (UGT1A1) i levern. Den vanligaste orsaken är att individen är homozygot för en insertion av två baser i promotorn för genen UGT1A1. Denna genvariant kallas (TA)7TAA  eller UGT1A1*28. GS leder till intermittent stegring av bilirubin vid infektioner, men bilirubinstegring kan förekomma även utan utlösande agens. GS kan också leda till bilirubinstegring vid viss läkemedelsbehandling. Irinotekan (Campto®) används vid metastaserande colorektal cancer och dess aktiva metabolit inaktiveras av UGT1A1. Det finns rapporter om att GS ger ökad risk för toxiska biverkningar av irinotekan. Tre metoder för att bestämma UGT1A1 har jämförts: PCR med elfores, PCR med smältpunktsanalys och PCR med fragmentanalys på sekvensator. Den sista metoden var bäst och användes för att genotypa UGT1A1 hos patienter med colorektal cancer från Nordic VI-studien. De behandlades med irinotekan i kombination med bolusinjektion eller infusion av 5-fluorouracil. Vi fann att  patienter med GS hade signifikant högre S-bilirubin före behandling jämfört med övriga patienter. De hade även ökad frekvens biverkningar av irinotekan (Fishers exakta test p=0,02). Genotypning av UGT1A1 kan således användas för att diagnostisera Gilberts syndrom hos patienter med oförklarad bilirubinstegring. Det kan även användas för att identifiera patienter med ökad risk för biverkningar av irinotekan.

Genetic factors account for about half of Alzheimers Disease (AD) risk and only about a quarter of that heritability is accounted for by known variants. Family based approaches to understanding AD genetics may be an effective way to identify additional risk factors. Here we report the results of whole exome sequencing (WES) and analyses done on pairs of AD affected cousins from 19 families from the Utah Population Database (UPDB) with a statistical excess of AD risk. WES variants passing quality control were additionally filtered by population frequency (minor allele <<> 0.01) and concordance between cousin pairs, resulting in 564 variants shared by at least one pair of cousins. For each of these variants we conducted in depth annotations using Ingenuity Variant Analysis (IVA), Wellderly Data Allele Frequencies, and literature searches. To further aid in variant prioritization we analyzed each variant for association with Age at Onset of AD, AD Risk, CSF AB42, CSF Tau, CSF PTau and Rate of Disease Decline in data from the Alzheimers Disease Genetics Consortium (ADGC) and from the Knight Alzheimers disease research center. Statistical analyses were conducted using PLINK. Twelve variants (rs201665195, rs28933981, rs148294193, rs147599881, rs61729902, rs140129800, rs191804178, rs200290640, rs199752248, rs45541434, rs141402160 and rs140914494) in eight genes (ABCA7, TTR, PELI3, FCHO1, SNAP91, COX6A2, MUC16, PIDD1, SYT5 and NOTCH3) were prioritized using a clear pipeline of IVA filters and the additional analysis information. We propose that these genes and variants are the most interesting for follow-up based on current knowledge.This family-based approach to finding rare AD variants adds to a growing body of research suggesting a role for NOTCH3 in late-onset AD. This approach replicated two known AD risk variants and also implicated novel putative risk AD variants and genes. These results suggest that further application of this method of using pairs of cousins may result in additional insights into AD genetics and the ability to find novel rare, causal AD variants.