The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate ...The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 10<sup>7</sup> FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.展开更多
Aim:To assess the efficacy and limitation of free/total prostate-specific antigen ratio(f/tPSA)at a single institution in Japan,focusing on the avoidance of pointless prostate biopsies.Methods:In total,631 men between...Aim:To assess the efficacy and limitation of free/total prostate-specific antigen ratio(f/tPSA)at a single institution in Japan,focusing on the avoidance of pointless prostate biopsies.Methods:In total,631 men between 44 and 93 years old(mean 69.8 years)with elevated PSA underwent power-Doppler ultrasoundgraphy-guided transrectal 10-core prostate biopsies at Niigata Cancer Center Hospital,and their histological features were investigated with total PSA (tPSA)and f/tPSA.Results:PCa was detected in 126 of 134 patients(94.3%)with tPSA of 26 ng/mL or higher.The detection rate was 59.4% for tPSA of 21-25 ng/mL,followed by 39.2% for 16-20 ng/mL,30.0% for 11-15 ng/mL, 20.0% for 4.1-10 ng/mL and 7.6% for≤4.0 ng/mL,f/tPSA of the PCa group was significantly lower than that of non-malignamt disorders in any tPSA ranges(mean 0.122 vs.0.160,P<0.001).Receiver-operating characteristics analyses showed that f/tPSA(AUC:0.664)performed more valuably than tPSA(AUC:0.559)in patients with tPSA between 3.0-10 ng/mL(P<0.01).Although f/tPSA of 0.250 for the cut-off value might miss 1.8% PCa patients,it potentially spares 9.2% of unnecessary biopsies.Conclusion:f/tPSA is more valuable compared with tPSA alone for the prediction of the occurrence of PCa.We recommend 0.250 as the cut-off value for f/tPSA in PCa screening for Asian men having so-called grey-zone tPSA.(Asian J Androl 2006 Jul;8:429-434)展开更多
文摘The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 10<sup>7</sup> FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.
文摘Aim:To assess the efficacy and limitation of free/total prostate-specific antigen ratio(f/tPSA)at a single institution in Japan,focusing on the avoidance of pointless prostate biopsies.Methods:In total,631 men between 44 and 93 years old(mean 69.8 years)with elevated PSA underwent power-Doppler ultrasoundgraphy-guided transrectal 10-core prostate biopsies at Niigata Cancer Center Hospital,and their histological features were investigated with total PSA (tPSA)and f/tPSA.Results:PCa was detected in 126 of 134 patients(94.3%)with tPSA of 26 ng/mL or higher.The detection rate was 59.4% for tPSA of 21-25 ng/mL,followed by 39.2% for 16-20 ng/mL,30.0% for 11-15 ng/mL, 20.0% for 4.1-10 ng/mL and 7.6% for≤4.0 ng/mL,f/tPSA of the PCa group was significantly lower than that of non-malignamt disorders in any tPSA ranges(mean 0.122 vs.0.160,P<0.001).Receiver-operating characteristics analyses showed that f/tPSA(AUC:0.664)performed more valuably than tPSA(AUC:0.559)in patients with tPSA between 3.0-10 ng/mL(P<0.01).Although f/tPSA of 0.250 for the cut-off value might miss 1.8% PCa patients,it potentially spares 9.2% of unnecessary biopsies.Conclusion:f/tPSA is more valuable compared with tPSA alone for the prediction of the occurrence of PCa.We recommend 0.250 as the cut-off value for f/tPSA in PCa screening for Asian men having so-called grey-zone tPSA.(Asian J Androl 2006 Jul;8:429-434)
