Viral antigen production in cell cultures on microcarriers Bovine parainfluenza 3 virus and MDBK cells.

Viral antigens can be obtained from infected mammalian cells cultivated on microcarriers. We have worked out parameters for the production of bovine parainfluenza 3 (PI-3) virus by Mandin-Darby Bovine Kidney (MDBK) cells cultivated on Cytodex 1 microcarriers (MCs) in spinners flasks and bioreactor using fetal bovine serum (FBS) supplemented Eagle minimal essential medium (Eagle-MEM). Medium renewal during the cell culture was shown to be crucial for optimal MCs loading (>90% MCs with confluent cell monolayers) and cell growth (2.5 x 10(6)cells/mL and a micro(x) (h(-1)) 0.05). Since cell cultures performed with lower amount of MCs (1g/L), showed good performances in terms of cell loading, we designed batch experiments with a lower concentration of MCs in view of optimizing the cell growth and virus production. Studies of cell growth with lower concentrations of MCs (0.85 g/L) showed that an increase in the initial cell seeding (from 7 to 40 cells/MC) led to a different kinetic of initial cell growth but to comparable final cell concentrations ((8-10)x10(5)cells/mL at 120 h) and cell loading (210-270 cells/MC). Upon infection with PI-3 virus, cultures showed a decrease in cell growth and MC loading directly related to the multiplicity of infection (moi) used for virus infection. Infected cultures showed also a higher consumption of glucose and production of lactate. The PI-3 virus and PI-3 antigen production among the cultures was not significantly different and attained values ranging from, respectively, 7-9 log(10) TCID(50)/mL and 1.5-2.2 OD. The kinetics of PI-3 virus production showed a sharp increase during the first 24h and those of PI-3 antigen increased after 24h. The differential kinetics of PI-3 virus and PI-3 antigen can be explained by the virus sensitivity to temperature. In view of establishing a protocol of virus production and based on the previous experiments, MDBK cell cultures performed under medium perfusion in a bioreactor of 1.2L were infected and the PI-3 virus production in 12L attained 12 log(10) TCID(50). Other than establishing a protocol for PI-3 production in MDBK cell cultures on Cytodex 1, the experiments are proposed as a basis for approaching the development of a virus production protocol in mammalian cells cultivated on microcarriers in bioreactors.

[Preoperative orientation: comprehensive analysis of the patient's point of view]. / Orientação pré-operatória: análise compreensiva sob a ótica do cliente.

In this study we approach the issue preoperatory orientation which we understand to be a vital practice in the field of Nursing also allowing the identification of same features at clients perception regarding the information achieved before operation. It is also suitable to recognize the failure of this procedure in the daily work at surgical units which upsets us with the absence of a guideline for preparing patients to surgical acts. Facing this need, we have decided to deeply investigate this issue to achieve the dimension, the description and analyzing the trajectory follow-up started in the first contact with the phenomenological approach. Thus, we make use of our patient operatory phase through his talk according to MARTINS & BICUDO's reference. So, this research aims at presenting the knowledge feature achieved through patients talk, its meaning and repercussion onto the individual plane as well as identifying what should alone improve it. That is the way phenomenological method questions the need of a preoperatory orientation which links us to theme dimension. In our experience of guiding patients to cope with their surgical intervention, we open a space that allows us to intervene under the perspective of conducting patients to recognize their existential dimension of experiencing an operation.