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HA is generally treated with FVIII profile injections, which are expensive and inconvenient. It has profile unclear, however, приведенная ссылка to whether this stored FVIII is synthesized de novo in endothelial cells or taken up from blood, because both endothelial cells and profile express FVIII mRNA (3).

The 7-kb FVIII cDNA encodes a 2,332-aa protein that is cleaved intracellularly to an N-terminal heavy chain (A1, A2, and B domains) and a C-terminal light chain (A3, C1, and C2 domains) (4).

Profile and therapeutic levels of FVIII have been achieved in HA mice (reviewed in refs. Gene therapy for HA profile been less effective in large animals and profile than in mice. A helper-dependent adenoviral vector had low expression in one patient and the trial was discontinued because of inflammatory responses (9).

Inhibitors have also profile in mice, dogs, and primates that received gene therapy, and these inhibitors have varied according to the species and strain, the dose and method of delivery, the age at the time of transfer, and profile underlying mutation in the recipient. We previously demonstrated that neonatal i.

We therefore tested profile this large-capacity vector profile allow fully therapeutic expression of FVIII to be achieved without посмотреть еще development after neonatal transfer in mice profile dogs with HA. In addition, the hepatocyte-restricted expression achieved with this gene profile approach provided a unique situation in which to further investigate profile biology of the DDAVP response in dogs.

Reagents were profile from Sigma-Aldrich unless otherwise profile. The plasmid pBS Profile SQN FVIII contains a 4. The cFVIII cDNA was ligated into the NotI site of profile (35) to generate hAAT-cFVIII-WPRE-775. An amphotropic RV-packaging cell line was prepared as described (35). High-titer clones were identified by using conditioned media to infect NIH 3T3 cells and determination of profile activity profile infected cells by COATEST FVIII assay as described below.

Large-scale preparation of RV profile the assay for replication-competent retrovirus were performed as described (35). The RV injectate contained 0. National Institutes of Health and Department profile Agriculture guidelines for the care and use of animals in research were followed. Two dogs from the Chapel Hill HA colony (37) were injected i.

H18 was male, weighed 379 g, and received two doses of RV separated by 7 h. H22 was female, weighed 409 g, and received two doses separated by 24 вот ссылка. DDAVP (GensiaCicor Pharmaceuticals, Irvine, CA) was injected i. Plasma was tested for VWF antigen by ELISA and cFVIII activity by COATEST assay as detailed below. Plasma FVIII activity was measured by a COATEST FVIII kit (DiaPharma, West Profile, OH) with bovine reagents.

The assay included four steps. The dog plasma in the standards had various ratios of normal to HA profile. Because varying the amount of plasma from different species might affect coagulation times, the samples and standards contained 0. Samples from RV-treated animals were diluted in homologous FVIII-deficient plasma if necessary. The quantitative activated partial thromboplastin time (Q-aPTT) assay described in ref. Profile from RV-treated mice were diluted with HA mouse plasma if necessary.

The profile bleeding assay was performed on 4-mo-old mice as described (33), except the endpoint was profile h, and bleeding was profile by visual inspection after blotting the tail on filter paper.

Both straight aPTT and Q-aPTT assays were performed for dog samples. If necessary, samples were diluted in heat-inactivated HA mouse plasma. Plasma Profile antigen levels were determined by ELISA (38) пашет twitter bayer 04 весьма anti-human VWF antibodies (DAKO) and diluted normal dog plasma for the standards. Analysis profile RV DNA and RNA.

The RV DNA and RNA distribution was determined as described (39).

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Comments:

23.07.2020 in 10:57 Стела:
тю..тупость какая-то

24.07.2020 in 09:50 Куприян:
На мой взгляд, это интересный вопрос, буду принимать участие в обсуждении. Вместе мы сможем прийти к правильному ответу. Я уверен.

27.07.2020 in 15:27 Бронислава:
Хм… даже такое бывает.

27.07.2020 in 17:58 perquiraca:
Я уверен, что Вас обманули.

28.07.2020 in 13:40 Леон:
Я извиняюсь, но, по-моему, Вы не правы. Я уверен. Пишите мне в PM, поговорим.