Despite 35 years of clinical trials, there has been little improvement in one-year survival rates with any chemotherapeutic regimen for the treatment of metastatic melanoma due to resistance to all known agents. Regardless of advances in detection and prevention, diagnosis of metastatic disease remains a death sentence. Resistance mechanisms, including aberrant kinase signaling and drug transport pumps, indicate a need for identification of other therapeutic targets that impinge upon multiple signaling pathways. The Abl family of non-receptor tyrosine kinases (c-Abl, Arg) has been indicted as a causative force in leukemia for more than three decades; however, their role in solid tumors has only recently been described. We first demonstrated that activated Abl family kinases promote breast cancer development and progression, and recently identified them to be novel therapeutic targets in metastatic melanoma cells by demonstrating that they promote proliferation, survival, invasion, and metastasis. We now present evidence that inhibitors of Abl family kinases abrogate resistance to a number of commonly used chemotherapeutics (i.e., 5-fluorouracil, cisplatin, paclitaxel, camptothecin) in a panel of breast cancer cells. We proceed to show that inhibitors of Abl family kinases, likewise, sensitize both breast cancer and melanoma cells to doxorubicin by blocking cell proliferation and dramatically inducing apoptosis. These findings were extended to advanced multi-drug resistant melanoma cells, in which we show for the first time that c- Abl promotes expression of the drug transporter, ABCB1, during acquired resistance, and drugs that inhibit c-Abl/Arg prevent ABCB1 expression and function. Moreover, c-Abl/Arg also promote acquired chemoresistance independent of ABCB1 by modulating multiple survival pathways. We demonstrate that c-Abl/Arg promote chemoresistance by upregulating STAT3, preventing doxorubicin-mediated conversion of NF-κB into a transcriptional repressor, activating an HSP27/p38/Akt survival pathway, and modulating ERK signaling. Therefore, c-Abl/Arg promote chemoresistance in highly resistant melanoma cells by impinging on drug transporter and cell survival pathways. Taken together, these data indicate that c-Abl/Arg inhibitors are likely to reverse acquired resistance in metastatic melanomas harboring activated c-Abl/Arg, and thus, may be effective in a combination regimen.

CBL proteins are E3 ubiquitin ligases and adaptor proteins. The mammalian homologs – CBL, CBL-B and CBL-3 show broad tissue expression; accordingly, the CBL proteins play roles in multiple cell types. We have investigated the function of the CBL-B protein in hematopoietic cells and fibroblasts. The causative agent of chronic myeloid leukemia (CML) is BCR-ABL. This oncogenic fusion down-modulates CBL-B protein levels, suggesting that CBL-B regulates either the development or progression of CML. To assess the involvement of CBL-B in CML, bone marrow transduction and transplantation (BMT) studies were performed. Recipients of BCR-ABL-infected CBL-B(-/-) cells succumbed to a CML-like myeloproliferative disease with a longer latency than the wild-type recipients. Peripheral blood white blood cell numbers were reduced, as were splenic weights. Yet despite the reduced leukemic burden, granulocyte numbers were amplified throughout the animals. As well, CBLB(-/-) bone marrow (BM) cells possessed defective BM homing capabilities. From these results we concluded that CBL-B negatively regulates granulopoiesis and that prolonged latency in our CBL-B(-/-) BMT animals was a function of perturbed homing.To develop an in vitro model to study CBL-B function we established mouse embryonic fibroblasts (MEFs) deficient in CBL-B expression. Transduction of the wild-type and CBL-B-deficient MEFs with BCR-ABL did not confer transformation; nevertheless, the role of CBL-B in fibroblasts was evaluated. The CBL-B(-/-) MEFs showed enhanced chemotactic migration toward serum in both Transwell migration and time-lapse video microscopy studies. The biochemical response to serum was extensively evaluated leading to the development of a model. We predict that CBL-B deficiency either: (a) augments GRB2-associated binding protein 2 (GAB2) phosphorylation leading to enhanced extracellular signal-regulated kinase (ERK) and protein kinase B (PKB / Akt) signaling, or (b) alleviates negative control of Vav3 resulting in stimulation of Rho effectors. In either case, our results reveal a negative regulatory role for CBL-B in fibroblast migration. The two studies detailed herein expand our knowledge of CBL-B function. They strongly suggest that CBL-B can modulate granulocyte proliferation and point toward a role for CBL-B in the motility of numerous cell types.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Reglerna vid tvångslikvidation innebär att styrelsen vid misstanke om kritisk kapitalbrist ska upprätta en kontrollbalansräkning. Uppvisar kontrollbalansräkningen en faktisk kapitalbrist ska en kontrollstämma hållas för att besluta om kapitalbristen ska läkas eller låta bolaget träda i likvidation. Underlåter styrelsen att upprätta en kontrollbalansräkning kan ett personligt betalningsansvar utdömas för styrelseledamöterna. Underlåtenheten omfattar även om styrelseledamöterna tror att de har upprättat en kontrollbalansräkning men den visar sig vara missvisande. Kontrollbalansräkningen kan i sådant fall anses vara felaktig, vilket i resulterar i att den inte är upprättad. Ett personligt betalningsansvar kan även aktualiseras för andra bolagsföreträdare förutsatt att de har vetskap om styrelsens underlåtenhet. Vid avvikelse från reglerna i 25 kap. ABL kan även ett skadeståndsansvar enligt 29 kap. ABL bli aktuellt. När borgenären åberopar personligt betalningsansvar, åligger det den att visa faktisk kapitalbrist samt en ansvarsgrund i 25 kap. ABL. Bolagsföreträdaren kan undkomma det personliga betalningsansvaret om den kan visa att ingen försumlighet har förelegat. Det finns således möjlighet att exculpera sig från ansvaret enligt 25 kap. ABL, medan någon motsvarande exculperingsregel inte finns för skadeståndsansvaret enligt 29 kap. ABL. Trots att grunder för exculpering existerar, tycks restriktivitet råda för bolagsföreträdaren att kunna exculpera sig från det personliga betalningsansvaret. HD har i NJA 2012 s. 858 lämnat vägledning angående försumlighetsprövningen i 25 kap. ABL. Avgörandet har även kommenterats av Stefan Lindskog och Håkan Andersson, varpå de har diskuterat hur försumlighetsprövningen ska bedömas. Upprättandet av en kontrollbalansräkning är till stor del en värderingsfråga beroende av redovisningsrättsliga normer, vilka inbegriper ett utrymme för alternativa lösningar. Konsekvenserna, av en diskutabel försumlighetsprövning och en värderingsproblematik, är att rättsläget för såväl ansvarsfrågan som exculperingsfrågan är svårbedömda.

Artificiell intelligens är digitala tekniker som har utvecklats snabbt de senaste åren. Allt fler uppgifter som tidigare krävt mänsklig färdighet och intelligens kan numera utföras autonomt av artificiell intelligens. Till följd av dess förmågor att på ett effektivt sätt behandla stora mängder information, lära sig nya saker, göra förutsägelser och föreslå handlingsalternativ samt i varierande grad handla efter egna uppfattningar, kan artificiell intelligens utgöra ett viktigt stöd vid beslutsfattande. Artificiell intelligens kan i teorin till och med ersätta människor. Syftet med uppsatsen är att undersöka under vilka förutsättningar artificiell intelligens kan användas för beslutsfattande i aktiebolag och att utvärdera hur svensk aktiebolagsrätt förhåller sig till den tekniska utveckling som sker på området. Frågor som uppstår är om artificiell intelligens kan ersätta aktieägare respektive styrelsen, i vilken utsträckning styrelsen kan delegera beslutsfattande till artificiell intelligens samt i vilken utsträckning styrelsen kan ta hjälp av artificiell intelligens vid sitt beslutsfattande. Även om vissa frågor med självklarhet kan besvaras, visar studien att det någonstans på vägen kommer en punkt där det inte går att avgöra hur artificiell intelligens ska behandlas. Undersökningen visar att artificiell intelligens varken kan ersätta aktieägare eller styrelsen i ett aktiebolag. Vad gäller styrelsens möjlighet att delegera beslutsfattande till artificiell intelligens samt om styrelsen kan ta hjälp av artificiell intelligens vid beslutsfattande, uppstår frågan om det kan anses oaktsamt av en styrelseledamot att använda artificiell intelligens på nämnt sätt och i sådant fall under vilka förutsättningar. En sådan aktsamhetsbedömning kan leda till att styrelseledamöter sanktioneras genom skadestånd. Rättsläget är oklart avseende om de uppgifter som styrelsen kan delegera till fysiska personer eller organ inom bolaget med ansvarsreducerande verkan, även kan delegeras till AI. Slutligen har styrelsen ett mycket stort handlingsutrymme att fatta beslut i aktiebolaget, under förutsättning att styrelsen iakttar bolagets intresse med omsorg. Det torde vara upp till styrelsen att avgöra om ett ärende kan beredas på ett tillfredsställande sätt med hjälp av AI.

A Leucemia Mieloide Crônica é uma doença hematopoiética associada a células estaminais que se manifesta principalmente com a expansão mielopoese. O cromossomo Philadelphia positivo PH+ é gerado por uma translocação recíproca (9, 22) (Q34, Q11) e pela fusão entre o gene Abel- son, essa fusáo codifica e desregula á protéına Tirosina Quinase, o suficiente para a iniciação e manutenção da doença. Inibidores como o Mesilato de Imatinibe revolucionou o tratamento de pacientes com leucemia mieloide crônica. As mutaç ões no domínio de quinase do Bcr-Abl, constituindo o mecanismo mais frequente de resistência adquirida para a terapia com inibidores da tirosina quinase. A mutação T315I atualmente e maior desafio para manutenção da Leucemia mieloide crônica na fase crônica, uma vez que os inibidores de Tirosina Quinase atualmente en- contrados no mercado são incapazes de mantê-la na forma controlada. Métodos computacionais baseados em fragmentos moleculares surgiram como uma nova estratégia para a descoberta de fármacos. Foi usado o programa LigBuilder para fazer geração das novas moléculas candidatas a ffármacos, usando dois métodos o Grow e Linker, as moléculas foram selecionadas por meio de docking com programa Glide da suite Maestro em cada mutação selecionadas da tirosina quinase, usando os melhores Gscores, para cada mutac¸a˜o, posteriormente foram submetidas ao programa QikProp, que tem a função comparar as moléculas com banco de dados de fármacos conhecidos. No último passo, é feito o estudo do docking das moléculas selecionas no sítio usando os protocolos de Induced Fit docking, que realiza o docking flexível-flexível, ou seja, ligante flexível e sítio de ligação da proteína flexível. / Chronic myeloid leukemia is associated with hematopoietic stem cell disorder that manifests itself primarily with myelopoiesis the expansion. The Philadelphia chromosome positive PH is generated by a reciprocal translocation (9, 22) (q34, q11), and by the merger of the Abelson gene fusion that encodes and deregulates quinase Tyrosine protein enough for the initiation and maintenance of disease. Tyrosine quinase provides a therapeutic target, which used to inhi- bition of this protein Known as tyrosine Quinase. Inhibitors such as Imatinibe mesylate has revolutionized the treatment of patients with chronic myeloid leukemia. Mutations in domain quinase Bcr-Abl, constituting the most frequent mechanism acquired resistance to therapy with tyrosine quinase. The T315I mutation and currently biggest challenge for maintenance of chro- nic myeloid leukemia in chronic phase, since inhibitors of tyrosine Quinase currently found on the market are unable to it maintaining it in a controlled manner, leading the patient achieved. Methods based on fragment docking emerged as a new strategy for drug discovery. evaluating all possible input locations and connecting the inhibitor and protein, and thus may provide a new molecule will be able to make effective inhibition. The docking studies are divided into three parts. At first, the fragments are placed to interact within the possible interaction sites. In the second step, the molecules are created from the best fragments which interacted with a particular website. In the last step, the study of molecules created in the docking site using the protocols of Induced Fit Docking which performs flexible, flexible docking ie flexible linker protein is made flexible.

Bakgrund och problem: Den förste juli 2009 genomfördes förändringarna i revisorsdirektivet och enligt de nya reglerna får en revisor enbart sägas upp i förtid av sin uppdragsgivare om det finns saklig grund till det. Bolagsverket är den organisation som granskar att underrättelse har kommit in.Förändringen är till för att stärka revisorns oberoende så att en klient inte kan entlediga revisorn utan saklig grund. Då återstår frågan vad är saklig grund och kan ett entledigande bero på att de yrkesetiska reglerna inte följs av revisorn?Då klienten måste ange en motivering kommer frågan kan motiveringen som anges påverka det förtroende som klienten har mot revisorn? Då allmänhetens förtroende för revisorns rykte har stor betydelse för hur revisionen i sig skapar trovärdighet i de finansiella rapporterna. Revisorns förtroende baseras i grunden på oberoende, etik och kompetens. Kan ryktet som revisorn har påverkas av ett negativt utlåtande eller påverkas det inte alls? Problemformuleringen blir därmed följande; vilka effekter ger förändringen gällande entledigandet av revisorer på deras förtroende?Syfte: Uppsatsen förmedlar en djupare förståelse för de effekter och reflektioner som uppkommit genom förändringen av revisorsdirektivet. En djupare förståelse för hur förtroendet har påverkats samt vad saklig grund innebär redovisas i studien. Även en förståelse för vad intressenter som revisorer, Bolagsverket och Revisorsnämnden har för åsikter redovisas. Studien ger också en förståelse för hur direktivet har anpassats till Sverige.Metod: Vi har använt oss av den kvalitativa metoden i uppsatsen och har intervjuat sex olika respondenter som arbetar inom revision, Bolagsverket och Revisorsnämnden. Respondenterna vi har intervjuat har varit informationsrika inom området och har medfört validitet till uppsatsen. Vi har genom intervjuerna fått en förståelse för hur de olika aktörerna ser på förändringen som har skett och vilka åsikter som förekommer.Resultat och slutsatser: Studiens resultat och slutsatser är att förtroendet inte har påverkats av lagändringen utan syftet är att stärka revisorns oberoende och hjälpa revisorn att inte bli entledigad utan orsak. Genom studien framkommer det genom respondenterna flera orsaker som kan tänkas vara saklig grund. Dock behöver begreppet preciseras och upprätta ett dokument som revisorer och företag kan följa.

A Leucemia Mielóide Crônica (LMC) é uma doença rara. No entanto, os avanços nas pesquisas básica e clínica nos últimos anos, colocaram a LMC em evidência sendo hoje uma neoplasia maligna potencialmente curável. O diagnóstico e tratamento desta doença são, no entanto, extremamente caros. Não havendo dados sistemáticos nem registros de incidência da LMC no Rio Grande do Sul ou no Brasil, o levantamento de dados baseado em registros dos centros de referência se justifica também para planejar ações em saúde. Entre 1996 e 2000, 276 casos foram diagnosticados. A estimativa de casos novos anuais foi de aproximadamente 0,6:100.000 habitantes, e a idade média no momento do diagnóstico foi 42 anos e 4 meses (±16 anos e 2 meses). Quanto ao tratamento e evolução destes pacientes, dos 257 avaliados, 56 (21,8%) foram submetidos ao transplante alogênico de medula óssea, com taxa de sobrevida em 5 anos de 75% e 27% para as fases crônica e acelerada/blástica, respectivamente. O tempo médio de sobrevida para os 257 pacientes foi de 47,7 meses (IC 43,3 - 52,1). Comparando ao relatado na literatura, encontramos um menor número anual de novos casos e também uma média de idade no diagnóstico mais baixa. Isto poderia ser explicado pela menor referência de idosos a serviços terciários de saúde. Para os pacientes transplantados, os resultados foram semelhantes aos relatados na literatura. / Although rare, the advances made in basic and clinical research throughout the last years have thrown a spotlight on CML. Diagnosis and treatment of CML is of high cost. Since there is no systematic data or information about the incidence of CML in Rio Grande do Sul or Brazil, the data obtained from reference centers serve to estimate the number of CML cases in our state to better plan health actions. Between 1996 and 2000, 276 cases were diagnosed. The annual estimate of new cases was approximately of 0,6:100,000 inhabitants, and the median age at diagnosis was 42 years and 4 months (±16 years and 2 months). The mean overall survival time for the 257 patients was 47,7 months (CI 43,3-52,1). That could be explained by the lack of referral for older patients. Regarding treatment and evolution, of the 257 valuable patients, 56 (21,8%) were submitted to allogeneic BMT with a five-year survival of 75% and 27% for chronic and accelerated/blastic phases, respectively. In conclusion, we found a lower estimated incidence and a lower median age at diagnosis. For the transplanted patients the results were similar to those reported in the literature.

Reagents that bind proteins are applicable in biology for detection of molecules, perturbation of signaling pathways and development of small-molecule pharmaceuticals. Protein ligands interact with proteins, inhibiting or altering their function. They are isolated from combinatorial libraries to interact with a specific target, using selection techniques such as phage display or yeast-two-hybrid assay. For the latter, one inconvenience is the detection of false positives, which can be solved by screening pools containing the samples to be tested, instead of individual samples. Samples are distributed in the pools following a pooling design. The PI-deconvolution pooling design was developed to screen cDNA libraries using the yeast-two-hybrid assay, which are smaller in size than protein ligand combinatorial libraries. Modifications to the PI-deconvolution screening technique were developed to adapt it to the screening of protein ligand combinatorial libraries using the yeast-two-hybrid assay. Every spot of the array containing the combinatorial library was randomly pooled. However, the yeast-two-hybrid assay loses sensitivity when strains are pooled. As PI-deconvolution requires detecting every interaction, we determined the optimal amount of library members that can be pooled in a spot, and the optimal number of replicates to ensure the detection of an interaction.<p> The yeast-two-hybrid assay was used to perform a screening of a combinatorial library with seven domains of BCR-ABL, which were pooled according to PI-deconvolution. BCR-ABL is a chimeric protein with unregulated kinase activity that is responsible for chronic myelogenous leukemia. The scaffold used in the combinatorial library was an engineered intein that forms lariat peptides. After a screening of this library was performed, positive interactions were detected in 775 spots of the arrays that contained 1432 positive hits. Only 53 spots were deconvoluted. The coding sequences of the lariat peptides were determined for 23 lariat peptides interacted with the GEF domain of BCR, and for ABL, two with the FABD domain, one with the SH1 domain, and one with the SH3 domain. Finally, a &beta;-galactosidase assay was performed to assess the affinity of the lariat peptides for their target.<p> The isolated lariat peptides are potential inhibitors of BCR-ABL that can have therapeutic potential. This study will improve other screenings of combinatorial libraries with the yeast-two-hybrid assay.

Reagents that bind proteins are applicable in biology for detection of molecules, perturbation of signaling pathways and development of small-molecule pharmaceuticals. Protein ligands interact with proteins, inhibiting or altering their function. They are isolated from combinatorial libraries to interact with a specific target, using selection techniques such as phage display or yeast-two-hybrid assay. For the latter, one inconvenience is the detection of false positives, which can be solved by screening pools containing the samples to be tested, instead of individual samples. Samples are distributed in the pools following a pooling design. The PI-deconvolution pooling design was developed to screen cDNA libraries using the yeast-two-hybrid assay, which are smaller in size than protein ligand combinatorial libraries. Modifications to the PI-deconvolution screening technique were developed to adapt it to the screening of protein ligand combinatorial libraries using the yeast-two-hybrid assay. Every spot of the array containing the combinatorial library was randomly pooled. However, the yeast-two-hybrid assay loses sensitivity when strains are pooled. As PI-deconvolution requires detecting every interaction, we determined the optimal amount of library members that can be pooled in a spot, and the optimal number of replicates to ensure the detection of an interaction.<p> The yeast-two-hybrid assay was used to perform a screening of a combinatorial library with seven domains of BCR-ABL, which were pooled according to PI-deconvolution. BCR-ABL is a chimeric protein with unregulated kinase activity that is responsible for chronic myelogenous leukemia. The scaffold used in the combinatorial library was an engineered intein that forms lariat peptides. After a screening of this library was performed, positive interactions were detected in 775 spots of the arrays that contained 1432 positive hits. Only 53 spots were deconvoluted. The coding sequences of the lariat peptides were determined for 23 lariat peptides interacted with the GEF domain of BCR, and for ABL, two with the FABD domain, one with the SH1 domain, and one with the SH3 domain. Finally, a &beta;-galactosidase assay was performed to assess the affinity of the lariat peptides for their target.<p> The isolated lariat peptides are potential inhibitors of BCR-ABL that can have therapeutic potential. This study will improve other screenings of combinatorial libraries with the yeast-two-hybrid assay.