ABSTRACT
Measles remains a major cause of worldwide infant mortality despite the use of current live attenuated vaccines. New approaches to measles virus (MV) vaccine development are critical to interrupt the spread of MV. In this study, we report the results using a DNA vaccine expressing a fusion of the measles hemagglutinin (H) protein and the complement component, C3d, to enhance the titers of neutralizing antibody. Plasmids were generated that expressed a secreted (s) form of H and the same form fused to three tandem copies of the murine homologue of C3d (sH-3C3d). Analysis of titers of the antibody raised in vaccinated mice indicated that immunizations with the DNA expressing sH-3C3d had higher titers of anti-H antibodies compared to serum from mice vaccinated with DNA expressing sH only. In addition, sH-3C3d elicited higher neutralizing antibody titers that inhibited MV induced plaque formation.
Subject(s)
Antibodies, Viral/immunology , Complement C3d/immunology , Hemagglutinins, Viral/immunology , Measles Vaccine/immunology , Measles virus/immunology , Animals , Cell Line , Chlorocebus aethiops , Complement C3d/genetics , Drug Evaluation, Preclinical , Genetic Vectors/genetics , Hemagglutinins, Viral/genetics , Humans , Immunization, Secondary , Kidney , Measles virus/genetics , Mice , Mice, Inbred BALB C , Neutralization Tests , Peptide Fragments/genetics , Peptide Fragments/immunology , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Transfection , Vaccination , Vaccines, DNA/immunology , Vero CellsABSTRACT
Steady state photoreceptor specific mRNA levels in bovine retina were studied during fetal maturation for five gene transcripts, including rhodopsin, arrestin (S-antigen), rod alpha-subunit of transducin, interphotoreceptor retinoid-binding protein (IRBP) and rod alpha-subunit of cGMP phosphodiesterase in order to understand mechanisms of gene regulation during photoreceptor development. A 10-15-fold increase in each transcript level begins between 5.5 and 6 months of gestation for each gene, suggesting a single coordinate induction event at this time. Quantitative analysis of transcriptional rates for each gene by nuclear run-on also reveals a coordinate increase at approximately the same time, demonstrating that induction is achieved by transcriptional activation. Interestingly, however, gene specific transcription rates in the pre-induction retina do not appear to parallel mRNA steady state levels. During fetal development neither the transcript level of each gene relative to the others nor relative mRNA turnover rates change substantially after the induction event at 5.5-6.0 months. However, at earlier times all genes exhibit higher mRNA turnover, implying that differential mRNA stability may also play an important role in determining steady-state levels.
Subject(s)
Embryonic and Fetal Development , Gene Expression Regulation , Photoreceptor Cells/metabolism , RNA, Messenger/biosynthesis , 3',5'-Cyclic-GMP Phosphodiesterases/biosynthesis , Animals , Antigens/biosynthesis , Arrestin , Blotting, Northern , Cattle , Eye Proteins/biosynthesis , Fetus/metabolism , Gestational Age , Membrane Proteins/biosynthesis , Retina/embryology , Retinol-Binding Proteins/biosynthesis , Rhodopsin , Transcription, Genetic , Transducin/biosynthesisABSTRACT
The in vitro activity of amifloxacin (WIN 49375), a new fluoroquinolone, was compared with the activities of antimicrobial agents that are commonly used for the treatment of urinary tract infection (cinoxacin, cephalexin, gentamicin, amoxicillin, trimethoprim, and trimethoprim-sulfamethoxazole) against 25 strains of Staphylococcus saprophyticus and 28 strains of Escherichia coli. Bacterial strains were isolated from urine specimens of college women with acute urinary tract infections. Bacterial isolates were more susceptible to trimethoprim-sulfamethoxazole, trimethoprim, and amifloxacin than to the other drugs tested. The in vitro activity of amifloxacin against S. saprophyticus had an inverse relation to increases in the pH of the test medium. Changes in the type of culture medium had no effect on the in vitro activity of amifloxacin. There was a direct relationship between increases in inoculum size and the MICs of amifloxacin.
