RESUMO
Background: Thermal stability [TS] is a part of the BCG vaccine characterization by which the consistency of process in BCG vaccine production could be confirmed. To enhance the TS of the vaccine, some prevalent stabilizers in different concentrations were added to the final formulation of BCG bulk prior to Freeze-drying process. We found a formulation more effective than the current stabilizer for retaining the higher viability of lyophilized BCG vaccine produced by Pasteur Institute of Iran
Methods: In the design of experiments using Taguchi method, lactose, trehalose, glucose, dextran, and monosodium glutamate were added to the final formulation of BCG bulk prior to freeze-drying process. Viability of the samples was determined by counting the colony forming unit
Results: Maximum signal-to-noise ratio equal to maximum TS and viability was obtained by adding lactose, dextran, and glutamate in defined concentrations
Conclusion: Adding the stabilizers had a significant impact on TS of BCG vaccine to meet the quality requirements
RESUMO
The ability to reducing death due to apoptosis and maintaining extensive levels on cell viability under serum free media in cell culture are important subjects in production of recombinant proteins. Insulin-like growth factor-I [IGF-I], is the growth factor of choice for mammalian cell proliferation in serum-free culture. In addition to its mitogenic activity, it has antiapoptotic activity protecting cultures from diverse death inducing stimuli. In this study, the effect of different concentrations of IGF-I examined on CHO-K1 [Chinese hamster ovary K1 cells] cell line for 24 and 48 hours. In this experimental study, the cell line was cultivated in DMEM supplemented with 10% fetal bovine serum [FBS]. Apoptosis process was induced in cells by methotrexate, serum was removed and then 10-50 ng/ml concentrations of IGF-I were added. Apoptosis was assessed by caspase 3 detection kit and cell proliferation and viability determined by MTT assay. Caspase 3 activity decreased significantly by increasing concentrations of IGF-I. In the highest concentration of IGF-I [50 ng/ml], 1.7 and 1.4 equal decreases of caspase 3 activities, compared with negative control group, were noted after 24 and 48 hours, respectively. It confirmed antiapoptotic activity of growth factor to maintain viability and protect cultures from apoptotic inducing stimuli [methotrexate]. IGF-I, as antiapoptotic factor, decreased programmed death of CHO-K1 cells in apoptotic induced conditions
RESUMO
Saffron [Zaaferan], botanical name Crocus sativus, is the most expensive spice in the world. It is derived from the dried stigma and pistil of the purple saffron crocus flowers. Iran is the largest saffron producer accounting for more than 80% of the world's production. Saffron contains an aeroallergen that causes reactive respiratory allergic reactions in atopic subjects. IgG antibody to allergens in the serum of allergic patients is not routinely measured. In this study in order to find out more about mechanism of allergy against saffron pollen, specific antibodies [IgE and IgG, total and subclasses] in atopic subjects were assayed. We used an ELISA assay for measuring specific IgE and IgG against saffron pollen extract in the sera of 38 atopic subjects [test group] and 20 non allergic subjects [control group]. The optical densities were compared between allergic subjects and non-allergic individuals. The prick test with saffron pollen extract was used to evaluate the cutaneous and specific antibody responses in the allergic subjects. The correlation was determined by statistical analysis. Specific saffron pollen IgE and IgG subclasses were found significantly higher in the allergic subjects than the control group. The immediate skin reaction was found positive in 70% of the test group. We report here, the existence of a positive correlation between specific IgE and skin reaction by prick test in atopic subjects [R=0.433]. A negative correlation between specific IgE and IgG4 subclass was also found [R=-0.576]. These data may be useful to understand the mechanism of allergy to saffron and may help in clarifying clinical manifestations and to prevent IgE production as well as therapeutic application