Estabilidade do fator de von Willebrand e fator VIII no crioprecipitado canino em diferentes protocolos de armazenamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A inquietude do homem segundo Agostinho: um estudo do tema nas Confissões, Livros I, VIII e X

Pós-graduação em Filosofia - FFC

Purification of coagulation factor VIII using chromatographic methods. Direct chromatography of plasma in anion exchange resins

Coagulation factor VIII (FVIII) concentrates are used in the treatment of patients with Hemophilia A. Human FVIII was purified directly from plasma using anion exchange chromatography followed by gel filtration. Three Q-Sepharose resins were tested, resulting in 40% recovery of FVIII activity using Q-Sepharose XL resin, about 80% using Q-Sepharose Fast Flow and 70% using the Q-Sepharose Big Beads. The vitamin K-dependent coagulation factors co-eluted with FVIII from the anion exchange columns. In the second step of purification, when Sepharose 6FF was used, 70% of FVIII activity was recovered free from vitamin K-dependent factors.

Production of human factor VIII-FL in 293T cells using the bicistronic MGMT(P140K)-retroviral vector

Hemophilia A is the most common X-linked bleeding disorder; it is caused by deficiency of coagulation factor VIII (FVIII). Replacement therapy with rFVIII produced from human cell line is a major goal for treating hemophilia patients. We prepared a full-length recombinant FVIII (FVIII-FL), using the pMFG-P140K retroviral vector. The IRES DNA fragment was cloned upstream to the P140K gene, providing a 9.34-kb bicistronic vector. FVIII-FL cDNA was then cloned upstream to IRES, resulting in a 16.6-kb construct. In parallel, an eGFP control vector was generated, resulting in a 10.1-kb construct. The 293T cells were transfected with these constructs, generating the 293T-FVIII-FL/P140K and 293T-eGFP/P140K cell lines. In 293T-FVIII-FL/P140K cells, FVIII and P140K mRNAs levels were 4,410 (+/- 931.7)- and 295,400 (+/- 75,769)-fold higher than in virgin cells. In 293T-eGFP/P140K cells, the eGFP and P140K mRNAs levels were 1,501,000 (+/- 493,700)- and 308,000 (+/- 139,300)-fold higher than in virgin cells. The amount of FVIII-FL was 0.2 IU/mL and 45 ng/mL FVIII cells or 4.4 IU/mu g protein. These data demonstrate the efficacy of the bicistronic retroviral vector expressing FVIII-FL and MGMT(P140K), showing that it could be used for producing the FVIII-FL protein in a human cell line.

SK-HEP cells and lentiviral vector for production of human recombinant factor VIII

Hemophilia A is caused by a deficiency in coagulation factor VIII. Recombinant factor VIII can be used as an alternative although it is unavailable for most patients. Here, we describe the production of a human recombinant B-domain-deleted FVIII (rBDDFVIII) by the human cell line SK-HEP-1, modified by a lentiviral vector rBDDFVIII was produced by recombinant SK-HEP cells (rSK-HEP) at 1.5-2.1 IU/10(6) in 24 h. The recombinant factor had increased in vitro stability when compared to commercial pdFVIII. The functionality of rBDDFVIII was shown by its biological activity and by tail-clip challenge in hemophilia A mice. The rSK-HEP cells grew in a scalable system and produced active rBDDFVIII, indicating that this platform production can be optimized to meet the commercial production scale needs.

Espacios de nadie y de todos: Territorio y sociedad en la frontera andalusí al norte del Tajo extremeño (Siglos VIII-XI)

[ES]Entre mediados del siglo VIII y mediados del siglo XII, el espacio comprendido entre el Sistema Central y el río Tajo en su parte extremeña ejerció como confín territorial de al- Andalus. A lo largo de todo este tiempo se sucedieron distintas fases en las que la frontera fluctuó según vinieran los aires cristianos del norte o islámicos del sur. Ello provocaría que estos territorios actuaran con relativa autonomía en muchos momentos de su historia andalusí y que se convirtieran en zonas donde confluían gentes de toda condición social y religiosa. En el presente trabajo tratamos de vislumbrar las dinámicas territoriales y sociales que se dieron en aquella región hasta finales del siglo XI, cuando se produjeron las conquistas cristianas de las ciudades islámicas más importantes de la frontera. [EN] The geographical area between the Spanish Central System and the part of the Tagus passing through Extremadura served as the boundary of the al-Andalus from the 8th to the 12th century. The boundary had different phases that fluctuated according to the influence exercised by the northern Christians or southern Muslims. This allowed these territories to have relative autonomy during its Andalusian history and turned them into zones where people from different social and religious background converged. The aim of this paper is to analyse the territorial and social dynamics of this area until the end of the 11th century, when the Christians conquered the most important Islamic towns of the borderland.

Paesaggi monastici nel Ravennate tra fonti scritte e dati archeologici (VIII – XIII secolo)

Il progetto di ricerca ha come oggetto di studio cinque monasteri ravennati (urbani e suburbani, maschili e femminili) di cui si volevano conoscere le strutture monastiche, le dotazioni patrimoniali (di cui è stata effettuata la mappatura) e le forme di economia sviluppate, utilizzando le fonti scritte provenienti dagli archivi monastici e i dati archeologici disponibili. Nello specifico, i monasteri selezionati (da intendersi come sede di monaci organizzati secondo una Regola) sono: Sant’Apollinare in Classe, San Severo, Santa Maria in cereseo, San Martino post ecclesiam maiorem, Sant’Andrea maggiore. L’arco cronologico preso in esame è compreso tra il secolo VIII e il XIII. Dall’elaborazione dei dati si è tentato di ricostruire la storia della comunità monastica così come è deducibile dalle fonti scritte, di conoscere le strutture dei monasteri e delle loro dipendenze, in base alle informazioni desumibili dalle fonti scritte, dagli scavi archeologici quando disponibili e dalle fonti iconografiche, di comprendere la formazione dei patrimoni monastici, la loro gestione e il ruolo svolto dai monasteri nel territorio in cui si concentravano i beni e di ricostruire il contesto naturale e le forme insediative in cui tali beni erano inseriti.