TREATMENT OF HEMOPHILIA WITH HUMAN FACTORIX PRODUCED IN MAMIMARY TISSUE OF TRANSGENIC MAMMALS

Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5\u27-untranslated and 3\u27-un-translated region, which is substituted with a 5- and 3\u27-end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described

TRANSGENIC NONHUMAN MAMMALS PRODUCING FIBRINOGEN IN MILKAND METHODS OF PRODUCING FIBRIN

A transgenic, non-human mammalian animal is capable of expressing a heterologous gene for human or other recombinant physiologically functional fibrinogen holoprotein or individual subunit chain polypeptides thereofora modified or fusion fibrinogen in mammary glands of the animals and secreting the expressed product into a body fluid. Methodol ogy employing such a mammal yields recombinant physiologically functional fibrinogens, subunit chain polypeptides thereof, and modified or fusion fibrinogens

TREATMENT OF HEMOPHILIA WITH HUMAN FACTORIX PRODUCED IN MAMIMARY TISSUE OF TRANSGENIC MAMMALS

Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5\u27-untranslated and 3\u27-un-translated region, which is substituted with a 5- and 3\u27-end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described

Affinity Purification of Biologically Active andInactive Forms of Recombinant Human Protein C Produced in Porcine Mammary Gland

Recombinant human protein C (rhPC) secreted in the milk of transgenic pigs was studied. \u27Ikansgenes having different regulatory elements of the murine milk protein, whey acidic protein, were used with cDNA and genomic human protein C (hPC) DNA sequences to obtain lower and higher expressing animals. The cDNA pigs had a range of expression of about 0.1-0.5 g/l milk. Two different genomic hPC pig lines have expressed 0.3 and 1-2 g/l, respectively. The rhPC was first purified at yields greater than 60 per cent using a monoclonal antibody (mAb) to the activation site on the heavy chain of hPC. Subsequent immunopurification with a calcium-dependent mAb directed to the y-carboxyglutamic acid domain of the light chain of hPC was used to fractionate a population having a higher specific anticoagulant activity in vW. The higher percentages of Ca2+-dependent conformers isolated from the total rhPC by immunopurification correlated well with higher specific activity and lower expression. A rate limitation in y-carboxylation of rhPC was clearly identified for the higher expressing animals. Thus, transgenic animals with high expression levels of complex recombinant proteins produced a lower percentage of biologically active protein

Transgenic non-human mammals expressing human coagulation factor VIII and von Willebrand factor

A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand\u27s Factor into milk may be used to stabilize newly-secreted Factor VIII

Hannah Arendt: El totalitarismo y sus horores (última parte)

_Seguna parte del ensayo sobre la obra de Hannah Arendt. El análisis filosófico-político realizado por Arendt marca indudablemente un hito en el camino del reconocimiento del totalitarismo como uno de los fenómenos más siniestro de la historia mundial, que, sin embargo, tiene raíces históricas y metafísicas que ponen en duda el carácter radical de las formas anteriores de la reflexión filosófica y política. La totalidad de las manifestaciones de la crueldad, arbitrariedad y burocratismo desalmado inherentes al poder totalitario superó los horizontes tradicionales del pensamiento filosófico, y el radicalismo del planteamiento del problema por parte de Hannah Arendt estimula de una manera nueva a evaluar y a comprender muchos acontecimientos y fenómenos de la civilización, la política y la cultura

Internationaler Kongress/Convegno internazionale Sprachkontakt und Mehrsprachigkeit. Zur Anglizismendiskussion in den Standardvariet\ue4ten des Deutschen und in Italien Contatto linguistico e plurilinguismo. La discussione sugli anglicismi nelle variet\ue0 standard del tedesco e in Italia

Il Dipartimento di Studi Interdisciplianri su Traduzione, Lingue e Culture (SITLeC) ha organizzato sotto la responsabilit\ue0 scientifica e organizzativa di Sandro M. Moraldo nella primavera del 2007 un convegno internazionale con il titolo 'Sprachkontakt und Mehrsprachigkeit. Zur Anglizismendiskussion in den Standardvariet\ue4ten des Deutschen und in Italien' // 'Contatto linguistico e plurilinguismo. La discussione sugli anglizismi nelle variet\ue0 standard del tedesco e in Italia.' Si trattava in un primo momento di tentare una ricostruzione di uno stato della questione dell\u2019influenza dell\u2019inglese sul tedesco e sull\u2019italiano. Ma non solo. La Svizzera era al centro del convegno. Emerge da ricerche sinora fatte che nelle scuole e nelle universit\ue0 svizzere aumenta la domanda di inglese che mette cos\uec a sua volta in discussione le seconde e terze lingue che sono spesso il francese e l\u2019italiano. Da questi dati emerge pertanto la preoccupazione che l\u2019inglese possa fungere da lingua ponte e pian piano mettere in crisi la convivenza tra le quattro lingue ufficiali nella Confederazione Elvetica. La domanda, alla quale i relatori hanno cercato di dare una risposta era proprio questa: Ll\u2019emergere dell\u2019inglese contribuisce a mettere in crisi il plurilinguismo, ossia il federalismo linguistico? Il continuo dilagare del fenomeno dei prestiti linguistici inglesi nella lingua tedesca e in quella italiana, specialmente dalla fine della seconda guerra mondiale, si presta a diverse considerazioni. Con il denominatore comune della forte presenza della lingua e della cultura angloamericana, il convegno intende riflettere una situazione molteplice che varia secondo le caratteristiche dei rapporti interculturali e delle pianificazioni politico-linguistiche ed economiche in atto

Cell-specific activity of the Human Immunodeficiency Virus enhancer repeat in vitro

The binding of nuclear proteins and the functional activity of the HIV-LTR enhancer repeats in different cell lines (Jurkat, CEM, H9, U937, Raji, B cells, T47D, HeLa, 293, and HepG2 cells) was investigated in vitro. Five distinct complexes formed with the enhancer repeat have been identified by an electrophoretic mobility shift assay. The distribution of these complexes varied qualitatively and quantitatively between nuclear proteins from different sources. In the extracts tested, transcription of the HIV-LTR 5′ deletion mutants (-453/80, -176/80, -117/80, -103/80, -65/80, and -48/80) was initiated correctly. Transcriptional stimulation dependent upon the presence of the enhancer repeat sequences was observed in all nuclear extracts and was highest in Jurkat, Raji, and B cell extracts. The presence of specific factors and the functional activity of the enhancer repeats as well as other regulatory units in a variety of cells indicates limited host-cell restriction of HIV transcription initiation in vitro