Šiuolaikinėje chemoterapijoje plačiai naudojami įvairūs chinonai. Pastaraisiais metais atkreiptas dėmesys į tai, kad chinonai, turintys aziridino grupes savo struktūroje yra labai toksiški piktybinėms ląstelėms. Šių junginių citotoksiškumo mechanizmai nėra pakankamai išanalizuoti. Šio darbo tikslas buvo ištirti skirtingų chinonų, tarp jų ir aziridino grupes turinčių, poveikį GLV – transformuotoms FLK ląstelėms. Tai yra avies embriono inkstų ląstelės, fibroblastų kilmės, transformuotos galvijų leukemijos virusu. Jos pasižymi vėžinėms ląstelėms būdingomis savybėmis: neribota proliferacija ir padidėjusiu prooksidantinių fermentų aktyvumu, kuris sąlygoja didesnį jautrumą chinonams, sukeliantiems oksidacinį stresą ir skatinantiems ląstelės žūtį. Gauti rezultatai parodė, kad chinonai, turintys savo struktūroje aziridino grupes (RH1 ir MeDZQ) yra toksiškesni FLK ląstelėms, negu chinonai, neturintys aziridino grupių (daunorubicinas ir durochinonas). Visų šių chinonų citotoksinį poveikį lemia kombinuotas mechanizmas: oksidacinis stresas, DT-diaforazės ir ksantinoksidazės veikla. Manoma, kad kai kurie aziridinilo grupes turintys chinonai gali būti labai perspektyviais priešvėžiniais vaistais. / Many of recently used chemotherapeutic agents have a quinoidal structure. It is supposed that the main mechanism of antitumour aziridinyl substituted benzoquinones is their two-electron reduction to alkylating products. However, other possible cytotoxicity mechanisms, e.g., oxidative stress, are studied insufficiently. The goal of our investigation was to analyse quinones influence to apoptosis in FLK (foetal lamb kidney fibroblasts) line, which posses increased prooxidant activities. The results show that quinones, which have aziridinyl substituents (RH1 and MeDZQ) are more cytotoxic than quinones without these substituents (daunorubicin and duroquinone). Cytotoxic effect of those quinones to cells is conditioned by combinative mechanism of oxidative stress, DT-diaforase and xantinoxidase action. It is supposed that aziridinyl substituted benzoquinones may be very perspective in antitumour treatment.

In the mouse embryo, the first definitive haematopoietic stem cells (HSCs), capable of repopulating adult irradiated mice, emerge at mid-gestation by embryonic day E11. At this stage, the aorta-gonad-mesonephros (AGM) region is able to initiate and expand HSCs. Recently, it has been shown that the development of HSC in the AGM region results from the maturation of haematopoietic precursors called pre-HSCs. Mounting evidence points at an endothelial origin for these cells, the haematogenic endothelium. Analysis of VEGFs mutants, a critical pathway for endothelial developement, suggested that it also plays a role during early haematopoiesis. The main receptor of the pathway, FLK-1 (also known as VEGRR2 or KDR), is expressed in early hematopoietic and endothelial cells in the mouse embryo. Knock-out mutants for Flk-1 showed a decrease of endothelial and intra-embryonic haematopoietic progenitors. Although Flk-1 has been identified as an essential gene for HSC emergence, its exact point of action in HSC development remains unknown. In this thesis, I investigated the role of FLK-1 signalling in haematopoietic development and defined precise stages and cell types during HSC emergence in which FLK-1 is critically involved. by using a reporter line and antibody staining, I demonstrated that FLK-1 is expressed in the pre-HSCs/HSC lineage. Germ-line Flk-1 knockout results in embryonic lethality at around E9.0, before HSC emergence, mainly due to defects in vasculogenesis. Since arterial specification precedes HSC formation, it has never been elucidated whether the haematopoietic defects found in the knockouts are a secondary effect of the loss of vasculature or it FLK-1 is directly involved in haematopoietic specification. Therefore, to determine the role of the receptor in HSC development, I used a conditional inducible mutagenesis approach that allowed the deletion of Flk-1 precisely when pre-HSCs mature into HSCs at E10.5 and E11.5. My data showed that Flk-1 deletion at these stages affects both endothelial and haematopoietic progenitors, as well as HSCs. This suggests that the VEGF pathway is not only essential in early stages of haematopoietic development, as previously demonstrated, but it may be also involved in the maturation of pre HSC into HSCs at later stages.

Made available in DSpace on 2014-06-11T19:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-11-04Bitstream added on 2014-06-13T20:50:57Z : No. of bitstreams: 1 belmonte_ea_me_jabo.pdf: 395445 bytes, checksum: 92da23e7783cda1b9d0618c6cb55f316 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Objetivou-se avaliar os parâmetros cardiovasculares e respiratórios, em cães anestesiados com isofluorano, submetidos à infusão contínua de morfina (3,3 μg/Kg/min) ou fentanil (0,03 μg/Kg/min), associados à lidocaína (50 μg/Kg/min) e cetamina (10 μg/Kg/min). Para tal, formaram-se dois grupos de oito animais, denominados morfina-lidocaína-cetamina MLK e fentanil-lidocaína-cetamina (FLK). Os cães foram pré-tratados com levomepromazina (0,5 mg/Kg IV), induzidos à anestesia com propofol (5 mg/Kg) e mantidos com isofluorano, ajustando-se a concentração para obterem-se valores de índice biespectral (BIS) entre 55 e 65. As mensurações da freqüência cardíaca (FC), duração (Ps) e amplitude (PmV) da onda P, intervalo PR, duração do complexo QRS, intervalo QT e RR, pressões arteriais sistólica (PAS), diastólica (PAD), e média (PAM), concentração de isofluorano expirado (EtISO), tensão de dióxido de carbono expirado (EtCO2), saturação de oxihemoglobina (SpO2) freqüência respiratória (f), tempo para extubação (Te), temperatura esofágica (T) e consumo de anestésico (ISOmL) tiveram início 30 minutos após a indução (M0), e após o início da infusão das soluções, em intervalos de 15 minutos (M15 a M75). Os dados foram submetidos à análise de perfil (P < 0,05) e ao Teste T (P < 0,05). À exceção da T (grupo FLK), não houve diferenças entre momentos para as demais variáveis. Entre grupos, EtISO (M30), FC (M0, M45, M60 e M75) e QRS (M60), ISOmL e T (M30, M45, M60 e M75), apresentaram médias do grupo MLK maiores que as do FLK. Para QT (M30 e M75), RR (M0, M60 e M75) e SpO2 (M60), as médias do grupo MLK foram menores que as do FLK. Concluiu-se que as soluções testadas não comprometem os parâmetros avaliados e não prejudicam a monitoração da profundidade anestésica por meio do BIS. / The aim of this study was investigate the effects over cardiovascular and respiratory system related with the continuous rate infusion of morphine (3,3mg/Kg/min) or fentanyl (0,03mg/Kg/min) associated with a solution of lidocaine (50ug/Kg/min) and ketamine (10ug/Kg/min). For such, two groups named morphine-lidocaine-ketamine (MLK) and fentanyl-lidocaine-ketamine (FLK) with eight dogs each were formed. As preanesthetic medication, the dogs received levomepromazine (0,5mg/Kg – IV). For induction and maintenance of anesthesia, propofol (5mg/Kg –IV) and isoflurane were used respectively. The values of bispectral index (BIS) were maintained between 55 and 65. The measurements of heart rate (HR), length (Ps) and amplitude (PmV) of the P wave, PR interval, duration of the QRS complex, QT interval and RR, systolic blood pressures (SBP), diastolic (DBP) and mean (MAP), expired concentration of isoflurane (EtISO), tension of carbon dioxide expired (ETCO2), oxyhemoglobin saturation (SpO2) respiratory rate (f), time to extubation (Te), esophageal temperature (T) and consumption of anesthetics (ISOmL) started 30 minutes after induction (M0), and after the infusion of solutions, at intervals of 15 minutes (M15 to M75). The data were submitted to analysis of profile (P <0.05) and the T test (P <0.05). With the exception of T (FLK group), there was no difference between moments for the other variables. Among groups, EtISO (M30), FC (M0, M45, M60 and M75) and QRS (M60), ISOmL and T (M30, M45, M60 and M75), showed means of MLK more than FLK. For QT (M30 and M75), RR (M0, M60 and M75) and SpO2 (M60), the means of MLK were smaller than FLK. The result of this study demonstrates that booth tested solutions not alters the respiratory or cardiovascular parameters, but, the solution with fentanyl promotes temperature reduction.

Orientador: Newton Nunes / Banca: João Moreira da Costa Neto / Banca: Paulo Sérgio Patto dos Santos / Resumo: Objetivou-se avaliar os parâmetros cardiovasculares e respiratórios, em cães anestesiados com isofluorano, submetidos à infusão contínua de morfina (3,3 μg/Kg/min) ou fentanil (0,03 μg/Kg/min), associados à lidocaína (50 μg/Kg/min) e cetamina (10 μg/Kg/min). Para tal, formaram-se dois grupos de oito animais, denominados morfina-lidocaína-cetamina MLK e fentanil-lidocaína-cetamina (FLK). Os cães foram pré-tratados com levomepromazina (0,5 mg/Kg IV), induzidos à anestesia com propofol (5 mg/Kg) e mantidos com isofluorano, ajustando-se a concentração para obterem-se valores de índice biespectral (BIS) entre 55 e 65. As mensurações da freqüência cardíaca (FC), duração (Ps) e amplitude (PmV) da onda P, intervalo PR, duração do complexo QRS, intervalo QT e RR, pressões arteriais sistólica (PAS), diastólica (PAD), e média (PAM), concentração de isofluorano expirado (EtISO), tensão de dióxido de carbono expirado (EtCO2), saturação de oxihemoglobina (SpO2) freqüência respiratória (f), tempo para extubação (Te), temperatura esofágica (T) e consumo de anestésico (ISOmL) tiveram início 30 minutos após a indução (M0), e após o início da infusão das soluções, em intervalos de 15 minutos (M15 a M75). Os dados foram submetidos à análise de perfil (P < 0,05) e ao Teste T (P < 0,05). À exceção da T (grupo FLK), não houve diferenças entre momentos para as demais variáveis. Entre grupos, EtISO (M30), FC (M0, M45, M60 e M75) e QRS (M60), ISOmL e T (M30, M45, M60 e M75), apresentaram médias do grupo MLK maiores que as do FLK. Para QT (M30 e M75), RR (M0, M60 e M75) e SpO2 (M60), as médias do grupo MLK foram menores que as do FLK. Concluiu-se que as soluções testadas não comprometem os parâmetros avaliados e não prejudicam a monitoração da profundidade anestésica por meio do BIS. / Abstract: The aim of this study was investigate the effects over cardiovascular and respiratory system related with the continuous rate infusion of morphine (3,3mg/Kg/min) or fentanyl (0,03mg/Kg/min) associated with a solution of lidocaine (50ug/Kg/min) and ketamine (10ug/Kg/min). For such, two groups named morphine-lidocaine-ketamine (MLK) and fentanyl-lidocaine-ketamine (FLK) with eight dogs each were formed. As preanesthetic medication, the dogs received levomepromazine (0,5mg/Kg - IV). For induction and maintenance of anesthesia, propofol (5mg/Kg -IV) and isoflurane were used respectively. The values of bispectral index (BIS) were maintained between 55 and 65. The measurements of heart rate (HR), length (Ps) and amplitude (PmV) of the P wave, PR interval, duration of the QRS complex, QT interval and RR, systolic blood pressures (SBP), diastolic (DBP) and mean (MAP), expired concentration of isoflurane (EtISO), tension of carbon dioxide expired (ETCO2), oxyhemoglobin saturation (SpO2) respiratory rate (f), time to extubation (Te), esophageal temperature (T) and consumption of anesthetics (ISOmL) started 30 minutes after induction (M0), and after the infusion of solutions, at intervals of 15 minutes (M15 to M75). The data were submitted to analysis of profile (P <0.05) and the T test (P <0.05). With the exception of T (FLK group), there was no difference between moments for the other variables. Among groups, EtISO (M30), FC (M0, M45, M60 and M75) and QRS (M60), ISOmL and T (M30, M45, M60 and M75), showed means of MLK more than FLK. For QT (M30 and M75), RR (M0, M60 and M75) and SpO2 (M60), the means of MLK were smaller than FLK. The result of this study demonstrates that booth tested solutions not alters the respiratory or cardiovascular parameters, but, the solution with fentanyl promotes temperature reduction. / Mestre

Detta examensarbete beskriver produktutveckling av en separeringsmaskin i ett inledande skede. Institutionen för Energi och Teknik vid SLU bedriver forskning inom fluglarvskompostering och behöver en utrustning för att separera larverna från behandlingsrester. Det finns ingen separationsutrustning direkt anpassad för detta ändamål därför behöver teknik utvärderas för att se vad som är lämpligt. Arbetet har utförts enligt TDB ”test-design-build” vilket innebär att det är praktisk produktutveckling i en verkstad där prototyper har tillverkats för att utvärdera olika separationstekniker. Testerna har lett till insikter som tagit arbetet vidare med en design som sedan blivit en mer raffinerad prototyp i pilotskala. Information har inhämtats löpande under projektet genom att konsultera olika professioner och genom sökningar i referensverk och bild- och videomaterial på internet. Egna studier på det material som skall separeras har bidragit med väsentlig kunskap för att lyckas med uppdraget. Varje prototyp innebar ett eget arbete i det stora projektet, med egna resultat och diskussioner. Resultaten från de initiala prototyperna är mer en funktionsbeskrivning medan den slutgiltiga prototypen har presenterats både beträffande funktion och med kvantitativ data. Resultaten visar på att separationen bör utföras med en kombinerad vibration- och vindsikt med kompletterande logistiklösningar för hantering av material in- och ur maskinen. Arbetet ger ett resultat i form av en prototyp tillika ett designförslag, som kan användas som en utgångspunkt i arbetet med att ta fram en optimerad produkt som kan separera FLK-materialet.

A new concept quickly gaining ground in the field of cancer research is that the inflammatory process plays a key role in cancer development and metastasis; however, the molecular mechanisms of such an involvement in cancer progression remain largely unspecified. YKL-40, also known as human cartilage glycoprotein 39, is a secreted heparin-binding protein with ties to both cancers and inflammatory disease. In these diseases, YKL-40 has been suggested to play a role in regulating tissue and extracellular matrix remodeling. It has been found that in certain cancers, including breast, colorectal and brain, that high YKL-40 serum levels correlate with poor outcome, and consequently it may serve as a biomarker. Our recent study has shown that tumor-derived YKL-40 acts as an angiogenic factor due to its ability to up-regulate vessel formation and metastasis during tumor development. However, blockade of the function of YKL-40, which implicates therapeutic value, has not been explored yet. The goal of this project was to better understand the importance of tumor-derived YKL-40 in angiogenesis through both functional and structural studies. By establishing a monoclonal YKL-40 antibody for blocking YKL-40, the function of tumor-derived YKL-40 in inducing endothelial cell angiogenesis and tumor cell survival was uncovered, confirming YKL-40's importance in tumor signaling as well as offering evidence in the benefit of its neutralization. Additionally, a postulated heparin-binding domain on YKL-40 was mutated in hopes of revealing the relevance of this binding ability on YKL-40's function and whether this could serve as a target in inhibiting YKL-40 signaling.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal. / L'utilisation d'oligodésoxyribonucléotides antisens est un outil puissant pour réguler 1'expression génique et le développement de nouvelles structures représente un domaine en pleine effervescence en chimie médicinale. En effet, nous cherchons à développer des molécules dont l'affinité de liaison pour l'ARNm serait plus élevée, tout en possédant la capacité d'activer la RNase H, l'enzyme qui reconnaît le complexe ADN / ARNm et qui dégrade l'ARNm. Il a déjà été démontré qu'une inversion de configuration en position C2' du ribose de l'ADN (ANA), combinée à la substitution du groupement 2'OH par un atome fluor (2'F-ANA) augmente l'affinité de liaison de l'oligonucléotide à l'ARNm. Lors de notre étude, nous avons évalué l'activité antisens d'oligonucléotides phosphorothioates chimériques composés de 2'-désoxy-2'-fluoro-P-D-arabinose et de 2'-désoxyribose (chimère S-2'F-ANA / ADN) comportant un sucre en confonnation L à leur extrémité 3'. Nous avons déterminé la capacité de ces molécules à inhiber 1'expression et la phosphorylation de Flk-1, un récepteur du VEGF ainsi que les effets médiés par le VEGF, soit la prolifération, la migration et la synthèse du «platelet-activating factor» (PAF) au niveau des cellules endothéliales (CE). Le traitement de CE d'aorte bovine avec les chimères S-2'F-ANA / ADN a pennis de réduire l'expression protéique ainsi que la phosphorylation de Flk-1 plus efficacement qu'un traitement avec les antisens d'ADN phosphorothioates (S-DNA) utilisés comme témoins positifs. Par contre, ces deux classes d'antisens ont inhibé les effets biologiques du VEGF de la même manière. Cette étude démontre aussi la capacité des antisens chimériques à activer la RNase H et leur meilleure affinité de liaison envers l'ARN comparativement aux antisens S-DNA.

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