It is the first time in Lithuania that practical application of semen filtration by Sephadex G-15 for the needs of artificial insemination industry was assessed. The efficacy of semen filtration was assessed applying a battery of sperm quality assays to study bovine spermatozoa quality before and after filtration, as well as after cryopreservation. The estimation of effect of different concentrations of progesterone, oestradiol, heparin separately or in combination, on different functional parameters of bull spermatozoa after thawing was performed. These are the first published studies applying integrated approach to study the effects of sperm challenge with several biologically active substances.

Made available in DSpace on 2014-12-17T14:03:41Z (GMT). No. of bitstreams: 1 TarcianaCGA.pdf: 634423 bytes, checksum: 54bd90fdc2abea42aaa8c6e698915963 (MD5) Previous issue date: 2006-07-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Fucans are a family of sulfated homo and teropolysaccharides respectively, composed mainly of a- (1?2) and a- (1?3) linked by L-fucose residues. Properties such as the ability to act as an anti-contraceptive, to reduce cholesterol levels, and to act as an anti-tumor agent are much related. We have focused our attention on the anticoagulant properties, platelet aggregation, hemorrhagic activity and complement system in vitro of commercial fucoidan (F) and their purified fractions (F1, F2 and F3) from Fucus vesiculosus obtained from fractionation of the fucoidan with different concentrations of acetone 1, 2 and 3v. These compounds were chemically characterized and the fucoidan (F) was modified by desulfation. The anticoagulant activity of the compounds was assessment by activated partial thromboplastin time (APTT) and prothrombine time assay (PT) using citrated normal human plasma. The results of APPT test showed that F, F1 and F2 have high anticoagulants activities 240.0 s (5 ?g). The F3 showed 73.7 s in the same concentrations. The results obtained with PT test to F, F1, F2 and F3 were 81.5 s, 120.0 s, 57.1 and 32.5 s respectively with 50 ?g. The dessulfated polymer showed a decrease in the anticoagulant activity in these two tests. Platelet aggregation assay was measured turbidimetrically with platelet aggregometer by method of Born. The aggregation platelet with F and fractions F1, F2 and F3 exhibited a two-phase answer in the concentration of 5 mg/mL with maximum aggregation of 76.36 ? 10.3% ; 69.54 ? 9.40%; 75.94 ? 9.01%; 51.13 ? 9.59% respectively. However, was observed a hipoaggregate profile F (15.17 ? 5.2%), F1 (7.40 ? 3.04 %), F2 (19.1 ? 5.41%) and F3 (5.09 ? 3.02%) at 0.1 mg/mL. The hemorrhagic activity assay was carried in Wistar rats and showed that these compounds have low hemorrhagic effect when compared to heparin. The complement system ( alternative pathway was made using non-sensibilized rabbit red blood cells The results of complement system essay showed that F , F2 and F3 have action inhibitory in relation to the group control 0.544, 0.697, 0.622 and 0.958 respectively The results showed that these compounds have action on this system. Interaction of the polisaccharides with proteins C3 and C4 showed that the fraction F1 stimulated the activity assay hemolytic using red blood cells / Fucanas s?o uma fam?lia de homo e hetero polissacar?deos sulfatados, formadas por uma cadeia central com liga??es α-(1→2) ou α-(1→3) unidas atrav?s de res?duos de L-fucose. Propriedades como a habilidade para agir como um anticoncepcional, reduzir n?veis de colesterol, e agir como um agente anti-tumoral foram relatadas. N?s focalizamos nossa aten??o nas propriedades anticoagulantes, agrega??o plaquet?ria, atividade anti-hemost?tica e sistema de complemento in vitro do fucoidan comercial e das fra??es F1, F2 e F3 de Fucus vesiculosus obtidas atrav?s do fracionamento do fucoidan por precipta??o com acetona (1, 2 e 3v). A atividade anticoagulante dos compostos foi avaliada pelo tempo de tromboplastina parcial ativado (APTT) e o tempo de protrombina (PT) usando plasma humano citratado. Os resultados do teste de APPT mostraram que o Fucoidan (F) e as fra??es F1 e F2 t?m altas atividades anticoagulantes 240,0 s (5 ?g), enquanto que a F3 mostrou 73,7s nas mesma concentra??o. Os resultados obtidos com PT para o F, F1, F2 e F3 foram 81,5 s, 120,0 s, 57,1 e 32,5 s respectivamente usando a massa de 50 ?g. A dessulfata??o do Fucoidan demonstrou uma diminui??o da atividade anticoagulante nos dois testes. O ensaio de agrega??o plaquet?ria foi realizado no agregometro de acordo com o m?todo de Born. A agrega??o plaquet?ria induzida pelo fucoidam e pelas fra??es de F1, F2 e F3 exibiram uma resposta bif?sica na concentra??o de 5 mg/mL com amplitude m?xima de agrega??o de 76,36% ? 10,3%; 69,54% ? 9,40%; 75,94% ? 9,01%; 51,13% ? 9,59% respectivamente. Por?m, na concentra??o 0,1 mg/mL foi observada um perfil hipoagregante para o Fucoidan (15,17% ? 5,2) e para as fra??es F1 (7,4% ? 3,04%), F2 (19,1% ? 5,41) e F3 (5,09% ? 3,02%). A atividade anti-hemost?tica foi realizada com ratos machos da linhagem Wistar e demonstrou que estes compostos t?m efeito hemorr?gico residual menor do que o da heparina. A a??o dos polissacar?deos fucosilados sulfatados na via alternativa do sistema complemento foi realizada atrav?s da utiliza??o de eritr?citos de coelhos n?o sensibilizados. Os resultados para o Sistema Complemento mostraram que o F, F2 e F3 t?m a??o neste sistema, apresentando efeito inibit?rio em rela??o ao grupo controle 0,544, 0,697, 0,622 e 0,958 respectivamente. Intera??es dos polissacar?deos com as prote?nas C3 e C4 demonstraram que a fra??o F1 estimula a atividade hemol?tica do complemento usando eritr?citos de coelho. Conclu?mos que estes a??cares t?m atividade anticoagulante e atuam como inibidores do sistema complemento sendo uma valiosa droga que pode ser empregada em doen?as relacionadas com inflama??o e coagula??o sang??nea